Human secreted proteins

ABSTRACT

The present invention relates to human secreted polypeptides, and isolated nucleic acid molecules encoding said polypeptides, useful for diagnosing and treating diseases, disorders, and/or conditions related to said human secreted proteins. Antibodies that bind these polypeptides are also encompassed by the present invention. Also encompassed by the invention are vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies. The invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The present invention further encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention.

FIELD OF THE INVENTION

The present invention relates to human secreted proteins/polypeptides, and isolated nucleic acid molecules encoding said proteins/polypeptides, useful for detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating diseases and disorders related to said proteins/polypeptides (relatedness may be by direct or indirect association, by cause, by consequence, or by effect on said diseases and disorders). Antibodies that bind these polypeptides are also encompassed by the present invention. Also encompassed by the invention are vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies. The invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The present invention further encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention.

BACKGROUND OF THE INVENTION

Unlike bacterium, which exist as a single compartment surrounded by a membrane, human cells and other eukaryotes are subdivided by membranes into many functionally distinct compartments. Each membrane-bounded compartment, or organelle, contains different proteins essential for the function of the organelle. The cell uses “sorting signals,” which are amino acid motifs located within the protein, to target proteins to particular cellular organelles.

One type of sorting signal, called a signal sequence, a signal peptide, or a leader sequence, directs a class of proteins to an organelle called the endoplasmic reticulum (ER). The ER separates the membrane-bounded proteins from all other types of proteins. Once localized to the ER, both groups of proteins can be further directed to another organelle called the Golgi apparatus. Here, the Golgi distributes the proteins to vesicles, including secretory vesicles, the cell membrane, lysosomes, and the other organelles.

Proteins targeted to the ER by a signal sequence can be released into the extracellular space as a secreted protein. For example, vesicles containing secreted proteins can fuse with the cell membrane and release their contents into the extracellular space—a process called exocytosis. Exocytosis can occur constitutively or after receipt of a triggering signal. In the latter case, the proteins are stored in secretory vesicles (or secretory granules) until exocytosis is triggered. Similarly, proteins residing on the cell membrane can also be secreted into the extracellular space by proteolytic cleavage of a “linker” holding the protein to the membrane.

Thus there exists a clear need for identifying and using novel secreted polynucleotides and polypeptides. Identification and sequencing of human genes is a major goal of modern scientific research. For example, by identifying genes and determining their sequences, scientists have been able to make large quantities of valuable human “gene products.” These include human insulin, interferon, Factor VIII, tumor necrosis factor, human growth hormone, tissue plasminogen activator, and numerous other compounds. Additionally, knowledge of gene sequences can provide the key to treatment or cure of genetic diseases (such as muscular dystrophy and cystic fibrosis).

SUMMARY OF THE INVENTION

The present invention relates to human secreted proteins/polypeptides, and isolated nucleic acid molecules encoding said proteins/polypeptides, useful for detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating diseases and disorders related to said proteins/polypeptides (relatedness may be by direct or indirect association, or by cause, consequence, or effect on said diseases and disorders). Antibodies that bind these polypeptides are also encompassed by the present invention. Also encompassed by the invention are vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies. The invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The present invention further encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention.

DETAILED DESCRIPTION

Polynucleotides and Polypeptides of the Invention

Description of Table 1A

Table 1A summarizes information concerning certain polynucleotides and polypeptides of the invention. The first column provides the gene number in the application for each clone identifier. The second column provides a unique clone identifier, “Clone ID:”, for a cDNA clone related to each contig sequence disclosed in Table 1A. Third column, the cDNA Clones identified in the second column were deposited as indicated in the third column (i.e. by ATCC Deposit No:Z and deposit date). Some of the deposits contain multiple different clones corresponding to the same gene. In the fourth column, “Vector” refers to the type of vector contained in the corresponding cDNA Clone identified in the second column. In the fifth column, the nucleotide sequence identified as “NT SEQ ID NO:X” was assembled from partially homologous (“overlapping”) sequences obtained from the corresponding cDNA clone identified in the second column and, in some cases, from additional related cDNA clones. The overlapping sequences were assembled into a single contiguous sequence of high redundancy (usually three to five overlapping sequences at each nucleotide position), resulting in a final sequence identified as SEQ ID NO:X. In the sixth column, “Total NT Seq.” refers to the total number of nucleotides in the contig sequence identified as SEQ ID NO:X.” The deposited clone may contain all or most of these sequences, reflected by the nucleotide position indicated as “5′ NT of Clone Seq.” (seventh column) and the “3′ NT of Clone Seq.” (eighth column) of SEQ ID NO:X. In the ninth column, the nucleotide position of SEQ ID NO:X of the putative start codon (methionine) is identified as “5′ NT of Start Codon.” Similarly, in column ten, the nucleotide position of SEQ ID NO:X of the predicted signal sequence is identified as “5′ NT of First AA of Signal Pep.” In the eleventh column, the translated amino acid sequence, beginning with the methionine, is identified as “AA SEQ ID NO:Y,” although other reading frames can also be routinely translated using known molecular biology techniques. The polypeptides produced by these alternative open reading frames are specifically contemplated by the present invention.

In the twelfth and thirteenth columns of Table 1A, the first and last amino acid position of SEQ ID NO:Y of the predicted signal peptide is identified as “First AA of Sig Pep” and “Last AA of Sig Pep.” In the fourteenth column, the predicted first amino acid position of SEQ ID NO:Y of the secreted portion is identified as “Predicted First AA of Secreted Portion”. The amino acid position of SEQ ID NO:Y of the last amino acid encoded by the open reading frame is identified in the fifteenth column as “Last AA of ORF”.

SEQ ID NO:X (where X may be any of the polynucleotide sequences disclosed in the sequence listing) and the translated SEQ ID NO:Y (where Y may be any of the polypeptide sequences disclosed in the sequence listing) are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below. For instance, SEQ ID NO:X is useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ID NO:X or the cDNA contained in the deposited clone. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling a variety of forensic and diagnostic methods of the invention. Similarly, polypeptides identified from SEQ ID NO:Y may be used, for example, to generate antibodies which bind specifically to proteins containing the polypeptides and the secreted proteins encoded by the cDNA clones identified in Table 1A and/or elsewhere herein

Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence. In these cases, the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).

Accordingly, for those applications requiring precision in the nucleotide sequence or the amino acid sequence, the present invention provides not only the generated nucleotide sequence identified as SEQ ID NO:X, and the predicted translated amino acid sequence identified as SEQ ID NO:Y, but also a sample of plasmid DNA containing a human cDNA of the invention deposited with the ATCC, as set forth in Table 1A. The nucleotide sequence of each deposited plasmid can readily be determined by sequencing the deposited plasmid in accordance with known methods

The predicted amino acid sequence can then be verified from such deposits. Moreover, the amino acid sequence of the protein encoded by a particular plasmid can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human cDNA, collecting the protein, and determining its sequence.

Also provided in Table 1A is the name of the vector which contains the cDNA plasmid. Each vector is routinely used in the art. The following additional information is provided for convenience.

Vectors Lambda Zap (U.S. Pat. Nos. 5,128,256 and 5,286,636), Uni-Zap XR (U.S. Pat. Nos. 5,128,256 and 5,286,636), Zap Express (U.S. Pat. Nos. 5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. et al., Nucleic Acids Res. 16:7583-7600 (1988); Alting-Mees, M. A. and Short, J. M., Nucleic Acids Res. 17:9494 (1989)) and pBK (Alting-Mees, M. A. et al., Strategies 5:58-61 (1992)) are commercially available from Stratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La Jolla, Calif., 92037. pBS contains an ampicillin resistance gene and pBK contains a neomycin resistance gene. Phagemid pBS may be excised from the Lambda Zap and Uni-Zap XR vectors, and phagemid pBK may be excised from the Zap Express vector. Both phagemids may be transformed into E. coli strain XL-1 Blue, also available from Stratagene

Vectors pSport1, pCMVSport 1.0, pCMVSport 2.0 and pCMVSport 3.0, were obtained from Life Technologies, Inc., P.O. Box 6009, Gaithersburg, Md. 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. See, for instance, Gruber, C. E., et al., Focus 15:59 (1993). Vector lafmid BA (Bento Soares, Columbia University, New York, N.Y.) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue. Vector pCR®2.1, which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, Calif. 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. See, for instance, Clark, J. M., Nuc. Acids Res. 16:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991).

The present invention also relates to the genes corresponding to SEQ ID NO:X, SEQ ID NO:Y, and/or a deposited cDNA (cDNA Clone ID). The corresponding gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include, but are not limited to, preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material.

Also provided in the present invention are allelic variants, orthologs, and/or species homologs. Procedures known in the art can be used to obtain full-length genes, allelic variants, splice variants, full-length coding portions, orthologs, and/or species homologs of genes corresponding to SEQ ID NO:X and SEQ ID NO:Y using information from the sequences disclosed herein or the clones deposited with the ATCC. For example, allelic variants and/or species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for allelic variants and/or the desired homologue.

The present invention provides a polynucleotide comprising, or alternatively consisting of, the nucleic acid sequence of SEQ ID NO:X and/or a cDNA contained in ATCC Deposit No.Z. The present invention also provides a polypeptide comprising, or alternatively, consisting of, the polypeptide sequence of SEQ ID NO:Y, a polypeptide encoded by SEQ ID NO:X, and/or a polypeptide encoded by a cDNA contained in ATCC deposit No.Z. Polynucleotides encoding a polypeptide comprising, or alternatively consisting of the polypeptide sequence of SEQ ID NO:Y, a polypeptide encoded by SEQ ID NO:X and/or a polypeptide encoded by the cDNA contained in ATCC Deposit No.Z, are also encompassed by the invention. The present invention further encompasses a polynucleotide comprising, or alternatively consisting of the complement of the nucleic acid sequence of SEQ ID NO:X, and/or the complement of the coding strand of the cDNA contained in ATCC Deposit No.Z.

Description of Table 1B (Comprised of Tables 1B.1 and 1B.2)

Table 1B.1 and Table 1B.2 summarize some of the polynucleotides encompassed by the invention (including cDNA clones related to the sequences (Clone ID:), contig sequences (contig identifier (Contig ID:) and contig nucleotide sequence identifiers (SEQ ID NO:X)) and further summarizes certain characteristics of these polynucleotides and the polypeptides encoded thereby. The first column of Tables 1B.1 and 1B.2 provide the gene numbers in the application for each clone identifier. The second column of Tables 1B.1 and 1B.2 provide unique clone identifiers, “Clone ID:”, for cDNA clones related to each contig sequence disclosed in Table 1A and/or Table 1B. The third column of Tables 1B.1 and 1B.2 provide unique contig identifiers, “Contig ID:” for each of the contig sequences disclosed in these tables. The fourth column of Tables 1B.1 and 1B.2 provide the sequence identifiers, “SEQ ID NO:X”, for each of the contig sequences disclosed in Table 1A and/or 1B.

Table 1B.1

The fifth column of Table 1B.1, “ORF (From-To)”, provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence of SEQ ID NO:X that delineates the preferred open reading frame (ORF) that encodes the amino acid sequence shown in the sequence listing and referenced in Table 1B.1 as SEQ ID NO:Y (column 6). Column 7 of Table 1B.1 lists residues comprising predicted epitopes contained in the polypeptides encoded by each of the preferred ORFs (SEQ ID NO:Y). Identification of potential immunogenic regions was performed according to the method of Jameson and Wolf (CABIOS, 4; 181-186 (1988)); specifically, the Genetics Computer Group (GCG) implementation of this algorithm, embodied in the program PEPTIDESTRUCTURE (Wisconsin Package v10.0, Genetics Computer Group (GCG), Madison, Wis.). This method returns a measure of the probability that a given residue is found on the surface of the protein. Regions where the antigenic index score is greater than 0.9 over at least 6 amino acids are indicated in Table 1B.1 as “Predicted Epitopes”. In particular embodiments, polypeptides of the invention comprise, or alternatively consist of, one, two, three, four, five or more of the predicted epitopes described in Table 1B.1. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly. Column 8 of Table 1B.1 (“Cytologic Band”) provides the chromosomal location of polynucleotides corresponding to SEQ ID NO:X. Chromosomal location was determined by finding exact matches to EST and cDNA sequences contained in the NCBI (National Center for Biotechnology Information) UniGene database. Given a presumptive chromosomal location, disease locus association was determined by comparison with the Morbid Map, derived from Online Mendelian Inheritance in Man (Online Mendelian Inheritance in Man, OMIM™. McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University (Baltimore, Md.) and National Center for Biotechnology Information, National Library of Medicine (Bethesda, Md.) 2000. World Wide Web URL: http://www.ncbi.nlm.nih.gov/omim/). If the putative chromosomal location of the Query overlaps with the chromosomal location of a Morbid Map entry, an OMIM identification number is disclosed in Table 1B.1, column 9 labeled “OMIM Disease Reference(s)”. A key to the OMIM reference identification numbers is provided in Table 5.

Table 1B.2

Column 5 of Table 1B.2, “Tissue Distribution” shows the expression profile of tissue, cells, and/or cell line libraries which express the polynucleotides of the invention. The first code number shown in Table 1B.2 column 5 (preceding the colon), represents the tissue/cell source identifier code corresponding to the key provided in Table 4. Expression of these polynucleotides was not observed in the other tissues and/or cell libraries tested. The second number in column 5 (following the colon), represents the number of times a sequence corresponding to the reference polynucleotide sequence (e.g., SEQ ID NO:X) was identified in the corresponding tissue/cell source. Those tissue/cell source identifier codes in which the first two letters are “AR” designate information generated using DNA array technology. Utilizing this technology, cDNAs were amplified by PCR and then transferred, in duplicate, onto the array. Gene expression was assayed through hybridization of first strand cDNA probes to the DNA array. cDNA probes were generated from total RNA extracted from a variety of different tissues and cell lines. Probe synthesis was performed in the presence of ³³P dCTP, using oligo(dT) to prime reverse transcription. After hybridization, high stringency washing conditions were employed to remove non-specific hybrids from the array. The remaining signal, emanating from each gene target, was measured using a Phosphorimager. Gene expression was reported as Phosphor Stimulating Luminescence (PSL) which rejects the level of phosphor signal generated from the probe hybridized to each of the gene targets represented on the array. A local background signal subtraction was performed before the total signal generated from each array was used to normalize gene expression between the different hybridizations. The value presented after “[array code]:” represents the mean of the duplicate values, following background subtraction and probe normalization. One of skill in the art could routinely use this information to identify normal and/or diseased tissue(s) which show a predominant expression pattern of the corresponding polynucleotide of the invention or to identify polynucleotides which show predominant and/or specific tissue and/or cell expression.

Description of Table 1C

Table 1C summarizes additional polynucleotides encompassed by the invention (including cDNA clones related to the sequences (Clone ID:), contig sequences (contig identifier (Contig ID:) contig nucleotide sequence identifiers (SEQ ID NO:X)), and genomic sequences (SEQ ID NO:B). The first column provides a unique clone identifier, “Clone ID:”, for a cDNA clone related to each contig sequence. The second column provides the sequence identifier, “SEQ ID NO:X”, for each contig sequence. The third column provides a unique contig identifier, “Contig ID:” for each contig sequence. The fourth column, provides a BAC identifier “BAC ID NO:A” for the BAC clone referenced in the corresponding row of the table. The fifth column provides the nucleotide sequence identifier, “SEQ ID NO:B” for a fragment of the BAC clone identified in column four of the corresponding row of the table. The sixth column, “Exon From-To”, provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence of SEQ ID NO:B which delineate certain polynucleotides of the invention that are also exemplary members of polynucleotide sequences that encode polypeptides of the invention (e.g., polypeptides containing amino acid sequences encoded by the polynucleotide sequences delineated in column six, and fragments and variants thereof).

Description of Table 1D

Table 1D: In preferred embodiments, the present invention encompasses a method of detecting, preventing, treating, and/or ameliorating a disease or disorder listed as listed in the “Preferred Indications” column of Table 1D (below); comprising administering to a patient (in which such detection, prevention, treatment, and/or amelioration is desired) a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) represented by Table 1A and Table 1D (in the same row as the disease or disorder to be treated is listed in the “Preferred Indications” column of Table 1D) in an amount effective to detect, prevent, treat, or ameliorate the disease or disorder.

As indicated in Table 1D, the polynucleotides, polypeptides, agonists, or antagonists of the present invention (including antibodies) can be used in assays to test for one or more biological activities. If these polynucleotides and polypeptides do exhibit activity in a particular assay, it is likely that these molecules may be involved in the diseases associated with the biological activity. Thus, the polynucleotides or polypeptides, or agonists or antagonists thereof (including antibodies) could be used to prevent, treat, or ameliorate the associated disease.

The present invention encompasses methods of detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating a disease or disorder. In preferred embodiments, the present invention encompasses a method of detecting, diagnosing, treating, preventing, or ameliorating a disease or disorder listed in the “Preferred Indications” column of Table 1D; comprising administering to a patient in which such treatment, prevention, or amelioration is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to treat, prevent, or ameliorate the disease or disorder. The first and second columns of Table 1D show the “Gene No.” and “cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may bc used in detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in Column 3 of Table 1D.

In another embodiment, the present invention also encompasses methods of preventing, treating, diagnosing, or ameliorating a disease or disorder listed in the “Preferred Indications” column of Table 1D; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in Column 3 of Table 1D.

The “Preferred Indication” column describes diseases, disorders, and/or conditions that may be treated, prevented, diagnosed, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The recitation of “Cancer” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., leukemias, cancers, and/or as described below under “Hyperproliferative Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D may be used for example, to diagnose, treat, prevent, and/or ameliorate a neoplasm located in a tissue selected from the group consisting of: colon, abdomen, bone, breast, digestive system, liver, pancreas, prostate, peritoneum, lung, blood (e.g., leukemia), endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid), uterus, eye, head and neck, nervous (central and peripheral), lymphatic system, pelvic, skin, soft tissue, spleen, thoracic, and urogenital.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a pre-neoplastic condition, selected from the group consisting of: hyperplasia (e.g., endometrial hyperplasia and/or as described in the section entitled “Hyperproliferative Disorders”), metaplasia (e.g., connective tissue metaplasia, atypical metaplasia, and/or as described in the section entitled “Hyperproliferative Disorders”), and/or dysplasia (e.g., cervical dysplasia, and bronchopulmonary dysplasia).

In another specific embodiment, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a benign dysproliferative disorder selected from the group consisting of: benign tumors, fibrocystic conditions, tissue hypertrophy, and/or as described in the section entitled “Hyperproliferative Disorders”.

The recitation of “Immune/Hematopoietic” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), blood disorders (e.g., as described below under “Immune Activity” “Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under “Infectious Disease”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having the “Immune/Hematopoietic” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: anemia, pancytopenia, leukopenia, thrombocytopenia, leukemias, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphocytic anemia (ALL), plasmacytomas, multiple myeloma, Burkitt's lymphoma, arthritis, asthma, AIDS, autoimmune disease, rheumatoid arthritis, granulomatous disease, immune deficiency, inflammatory bowel disease, sepsis, neutropenia, neutrophilia, psoriasis, immune reactions to transplanted organs and tissues, systemic lupus erythematosis, hemophilia, hypercoagulation, diabetes mellitus, endocarditis, meningitis, Lyme Disease, and allergies.

The recitation of “Reproductive” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the reproductive system (e.g., as described below under “Reproductive System Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Reproductive” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cryptorchism, prostatitis, inguinal hernia, varicocele, leydig cell tumors, verrucous carcinoma, prostatitis, malacoplakia, Peyronie's disease, penile carcinoma, squamous cell hyperplasia, dysmenorrhea, ovarian adenocarcinoma, Turner's syndrome, mucopurulent cervicitis, Sertoli-leydig tumors, ovarian cancer, uterine cancer, pelvic inflammatory disease, testicular cancer, prostate cancer, Klinefelter's syndrome, Young's syndrome, premature ejaculation, diabetes mellitus, cystic fibrosis, Kartagener's syndrome, testicular atrophy, testicular feminization, anorchia, ectopic testis, epididymitis, orchitis, gonorrhea, syphilis, testicular torsion, vasitis nodosa, germ cell tumors, stromal tumors, dysmenorrhea, retroverted uterus, endometriosis, fibroids, adenomyosis, anovulatory bleeding, amenorrhea, Cushing's syndrome, hydatidiform moles, Asherman's syndrome, premature menopause, precocious puberty, uterine polyps, dysfunctional uterine bleeding, cervicitis, chronic cervicitis, mucopurulent cervicitis, cervical dysplasia, cervical polyps, Nabothian cysts, cervical erosion, cervical incompetence, cervical neoplasms, pseudohermaphroditism, and premenstrual syndrome.

The recitation of “Musculoskeletal” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the immune system (e.g., as described below under “Immune Activity”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Musculoskeletal” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: bone cancers (e.g., osteochondromas, benign chondromas, chondroblastoma, chondromyxoid fibromas, osteoid osteomas, giant cell tumors, multiple myeloma, osteosarcomas), Paget's Disease, rheumatoid arthritis, systemic lupus erythematosus, osteomyelitis, Lyme Disease, gout, bursitis, tendonitis, osteoporosis, osteoarthritis, muscular dystrophy, mitochondrial myopathy, cachexia, and multiple sclerosis.

The recitation of “Cardiovascular” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the cardiovascular system (e.g., as described below under “Cardiovascular Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cardiovascular” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: myxomas, fibromas, rhabdomyomas, cardiovascular abnormalities (e.g., congenital heart defects, cerebral arteriovenous malformations, septal defects), heart disease (e.g., heart failure, congestive heart disease, arrhythmia, tachycardia, fibrillation, pericardial Disease, endocarditis), cardiac arrest, heart valve disease (e.g., stenosis, regurgitation, prolapse), vascular disease (e.g., hypertension, coronary artery disease, angina, aneurysm, arteriosclerosis, peripheral vascular disease), hyponatremia, hypernatremia, hypokalemia, and hyperkalemia.

The recitation of “Mixed Fetal” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Mixed Fetal” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: spina bifida, hydranencephaly, neurofibromatosis, fetal alcohol syndrome, diabetes mellitus, PKU, Down's syndrome, Patau syndrome, Edwards syndrome, Turner syndrome, Apert syndrome, Carpenter syndrome, Conradi syndrome, Crouzon syndrome, cutis laxa, Cornelia de Lange syndrome, Ellis-van Creveld syndrome, Holt-Oram syndrome, Kartagener syndrome, Meckel-Gruber syndrome, Noonan syndrome, Pallister-Hall syndrome, Rubinstein-Taybi syndrome, Scimitar syndrome, Smith-Lemli-Opitz syndrome, thromocytopenia-absent radius (TAR) syndrome, Treacher Collins syndrome, Williams syndrome, Hirschsprung's disease, Meckel's diverticulum, polycystic kidney disease, Turner's syndrome, and gonadal dysgenesis, Klippel-Feil syndrome, Ostogenesis imperfecta, muscular dystrophy, Tay-Sachs disease, Wilm's tumor, neuroblastoma, and retinoblastoma.

The recitation of “Excretory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and renal disorders (e.g., as described below under “Renal Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Excretory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: bladder cancer, prostate cancer, benign prostatic hyperplasia, bladder disorders (e.g., urinary incontinence, urinary retention, urinary obstruction, urinary tract Infections, interstitial cystitis, prostatitis, neurogenic bladder, hematuria), renal disorders (e.g., hydronephrosis, proteinuria, renal failure, pyelonephritis, urolithiasis, reflux nephropathy, and unilateral obstructive uropathy).

The recitation of “Neural/Sensory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the nervous system (e.g., as described below under “Neural Activity and Neurological Diseases”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Neural/Sensory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: brain cancer (e.g., brain stem glioma, brain tumors, central nervous system (Primary) lymphoma, central nervous system lymphoma, cerebellar astrocytoma, and cerebral astrocytoma, neurodegenerative disorders (e.g., Alzheimer's Disease, Creutzfeldt-Jakob Disease, Parkinson's Disease, and Idiopathic Presenile Dementia), encephalomyelitis, cerebral malaria, meningitis, metabolic brain diseases (e.g., phenylketonuria and pyruvate carboxylase deficiency), cerebellar ataxia, ataxia telangiectasia, and AIDS Dementia Complex, schizophrenia, attention deficit disorder, hyperactive attention deficit disorder, autism, and obsessive compulsive disorders.

The recitation of “Respiratory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the respiratory system (e.g., as described below under “Respiratory Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Respiratory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cancers of the respiratory system such as larynx cancer, pharynx cancer, trachea cancer, epiglottis cancer, lung cancer, squamous cell carcinomas, small cell (oat cell) carcinomas, large cell carcinomas, and adenocarcinomas. Allergic reactions, cystic fibrosis, sarcoidosis, histiocytosis X, infiltrative lung diseases (e.g., pulmonary fibrosis and lymphoid interstitial pneumonia), obstructive airway diseases (e.g., asthma, emphysema, chronic or acute bronchitis), occupational lung diseases (e.g., silicosis and asbestosis), pneumonia, and pleurisy.

The recitation of “Endocrine” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the respiratory system (e.g., as described below under “Respiratory Disorders”), renal disorders (e.g., as described below under “Renal Disorders”), and disorders of the endocrine system (e.g., as described below under “Endocrine Disorders”.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having an “Endocrine” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cancers of endocrine tissues and organs (e.g., cancers of the hypothalamus, pituitary gland, thyroid gland, parathyroid glands, pancreas, adrenal glands, ovaries, and testes), diabetes (e.g., diabetes insipidus, type I and type II diabetes mellitus), obesity, disorders related to pituitary glands (e.g., hyperpituitarism, hypopituitarism, and pituitary dwarfism), hypothyroidism, hyperthyroidism, goiter, reproductive disorders (e.g. male and female infertility), disorders related to adrenal glands (e.g., Addison's Disease, corticosteroid deficiency, and Cushing's Syndrome), kidney cancer (e.g., hypernephroma, transitional cell cancer, and Wilm's tumor), diabetic nephropathy, interstitial nephritis, polycystic kidney disease, glomerulonephritis (e.g., IgM mesangial proliferative glomerulonephritis and glomerulonephritis caused by autoimmune disorders; such as Goodpasture's syndrome), and nephrocalcinosis.

The recitation of “Digestive” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the gastrointestinal system (e.g., as described below under “Gastrointestinal Disorders”.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Digestive” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: ulcerative colitis, appendicitis, Crohn's disease, hepatitis, hepatic encephalopathy, portal hypertension, cholelithiasis, cancer of the digestive system (e.g., biliary tract cancer, stomach cancer, colon cancer, gastric cancer, pancreatic cancer, cancer of the bile duct, tumors of the colon (e.g., polyps or cancers), and cirrhosis), pancreatitis, ulcerative disease, pyloric stenosis, gastroenteritis, gastritis, gastric atropy, benign tumors of the duodenum, distension, irritable bowel syndrome, malabsorption, congenital disorders of the small intestine, bacterial and parasitic infection, megacolon, Hirschsprung's disease, aganglionic megacolon, acquired megacolon, colitis, anorectal disorders (e.g., anal fistulas, hemorrhoids), congenital disorders of the liver (e.g., Wilson's disease, hemochromatosis, cystic fibrosis, biliary atresia, and alpha1-antitrypsin deficiency), portal hypertension, cholelithiasis, and jaundice.

The recitation of “Connective/Epithelial” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), cellular and genetic abnormalities (e.g., as described below under “Diseases at the Cellular Level”), angiogenesis (e.g., as described below under “Anti-Angiogenesis Activity”), and or to promote or inhibit regeneration (e.g., as described below under “Regeneration”), and wound healing (e.g., as described below under “Wound Healing and Epithelial Cell Proliferation”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Connective/Epithelial” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: connective tissue metaplasia, mixed connective tissue disease, focal epithelial hyperplasia, epithelial metaplasia, mucoepithelial dysplasia, graft v. host disease, polymyositis, cystic hyperplasia, cerebral dysplasia, tissue hypertrophy, Alzheimer's disease, lymphoproliferative disorder, Waldenstron's macroglobulinemia, Crohn's disease, pernicious anemia, idiopathic Addison's disease, glomerulonephritis, bullous pemphigoid, Sjogren's syndrome, diabetes mellitus, cystic fibrosis, osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, osteoporosis, osteocarthritis, periodontal disease, wound healing, relapsing polychondritis, vasculitis, polyarteritis nodosa, Wegener's granulomatosis, cellulitis, rheumatoid arthritis, psoriatic arthritis, discoid lupus erythematosus, systemic lupus erythematosus, scleroderma, CREST syndrome, Sjogren's syndrome, polymyositis, dermatomyositis, mixed connective tissue disease, relapsing polychondritis, vasculitis, Henoch-Schonlein syndrome, erythema nodosum, polyarteritis nodosa, temporal (giant cell) arteritis, Takayasu's arteritis, Wegener's granulomatosis, Reiter's syndrome, Behcet's syndrome, ankylosing spondylitis, cellulitis, keloids, Ehler Danlos syndrome, Marfan syndrome, pseudoxantoma elasticum, osteogenese imperfecta, chondrodysplasias, epidermolysis bullosa, Alport syndrome, and cutis laxa.

Description of Table 1E

Table 1E provides information related to biological activities and preferred indications for polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof). Table 1E also provides information related to assays which may be used to test polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) for the corresponding biological activities. The first column (“Gene No.”) provides the gene number in the application for each clone identifier. The second column (“cDNA Clone ID:”) provides the unique clone identifier for each clone as previously described and indicated in Tables 1A, 1B, 1C, and 1D. The third column (“AA SEQ ID NO:Y”) indicates the Sequence Listing SEQ ID Number for polypeptide sequences encoded by the corresponding cDNA clones (also as indicated in Tables 1A, 1B, and 2). The fourth column (“Biological Activity”) indicates a biological activity corresponding to the indicated polypeptides (or polynucleotides encoding said polypeptides). The fifth column (“Exemplary Activity Assay”) further describes the corresponding biological activity and provides information pertaining to the various types of assays which may be performed to test, demonstrate, or quantify the corresponding biological activity. The sixth column (“Preferred Indications”) describes particular embodiments of the invention and indications (e.g. pathologies, diseases, disorders, abnormalities, etc.) for which polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) may be used in detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating.

Table 1E describes the use of FMAT technology, inter alia, for testing or demonstrating various biological activities. Fluorometric microvolume assay technology (FMAT) is a fluorescence-based system which provides a means to perform nonradioactive cell- and bead-based assays to detect activation of cell signal transduction pathways. This technology was designed specifically for ligand binding and immunological assays. Using this technology, fluorescent cells or beads at the bottom of the well are detected as localized areas of concentrated fluorescence using a data processing system. Unbound flurophore comprising the background signal is ignored, allowing for a wide variety of homogeneous assays. FMAT technology may be used for peptide ligand binding assays, immunofluorescence, apoptosis, cytotoxicity, and bead-based immunocapture assays. See, Miraglia S et. al., “Homogeneous cell and bead based assays for highthroughput screening using flourometric microvolume assay technology,” Journal of Biomolecular Screening; 4:193-204 (1999). In particular, FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides (including polypeptide fragments and variants) to activate signal transduction pathways. For example, FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides to upregulate production of immunomodulatory proteins (such as, for example, interleukins, GM-CSF, Rantes, and Tumor Necrosis factors, as well as other cellular regulators (e.g. insulin)).

Table 1E also describes the use of kinase assays for testing, demonstrating, or quantifying biological activity. In this regard, the phosphorylation and de-phosphorylation of specific amino acid residues (e.g. Tyrosine, Scrine, Thrconinc) on cell-signal transduction proteins provides a fast, reversible means for activation and de-activation of cellular signal transduction pathways. Moreover, cell signal transduction via phosphorylation/de-phosphorylation is crucial to the regulation of a wide variety of cellular processes (e.g. proliferation, differentiation, migration, apoptosis, etc.). Accordingly, kinase assays provide a powerful tool useful for testing, confirming, and/or identifying polypeptides (including polypeptide fragments and variants) that mediate cell signal transduction events via protein phosphorylation. See e.g., Forrer, P., Tamaskovic R., and Jaussi, R. “Enzyme-Linked Immunosorbent Assay for Measurement of JNK, ERK, and p38 Kinase Activities” Biol. Chem. 379(8-9): 1101-1110 (1998).

Description of Table 1F

Polynucleotides encoding polypeptides of the present invention can be used in assays to test for one or more biological activities. One such biological activity which may be tested includes the ability of polynucleotides and polypeptides of the invention to stimulate up-regulation or down-regulation of expression of particular genes and proteins. Hence, if polynucleotides and polypeptides of the present invention exhibit activity in altering particular gene and protein expression patterns, it is likely that these polynucleotides and polypeptides of the present invention may be involved in, or capable of effecting changes in, diseases associated with the altered gene and protein expression profiles. Hence, polynucleotides, polypeptides, or antibodies of the present invention could be used to treat said associated diseases.

TaqMan® assays may be performed to assess the ability of polynucleotides (and polypeptides they encode) to alter the expression pattern of particular “target” genes. TaqMan® reactions are performed to evaluate the ability of a test agent to induce or repress expression of specific genes in different cell types. TaqMan® gene expression quantification assays (“TaqMan® assays”) are well known to, and routinely performed by, those of ordinary skill in the art. TaqMan® assays are performed in a two step reverse transcription/polymerase chain reaction (RT-PCR). In the first (RT) step, cDNA is reverse transcribed from total RNA samples using random hexamer primers. In the second (PCR) step, PCR products are synthesized from the cDNA using gene specific primers.

To quantify gene expression the Taqman® PCR reaction exploits the 5′ nuclease activity of AmpliTaq Gold® DNA Polymerase to cleave a Taqman® probe (distinct from the primers) during PCR. The Taqman® probe contains a reporter dye at the 5′-end of the probe and a quencher dye at the 3′ end of the probe. When the probe is intact, the proximity of the reporter dye to the quencher dye results in suppression of the reporter fluorescence. During PCR, if the target of interest is present, the probe specifically anneals between the forward and reverse primer sites. AmpliTaq Fold DNA Polymerase then cleaves the probe between the reporter and quencher when the probe hybridizes to the target, resulting in increased fluorescence of the reporter (see FIG. 2). Accumulation of PCR products is detected directly by monitoring the increase in fluorescence of the reporter dye.

After the probe fragments are displaced from the target, polymerization of the strand continues. The 3′-end of the probe is blocked to prevent extension of the probe during PCR. This process occurs in every cycle and does not interfere with the exponential accumulation of product. The increase in fluorescence signal is detected only if the target sequence is complementary to the probe and is amplified during PCR. Because of these requirements, any nonspecific amplification is not detected.

For test sample preparation, vector controls or constructs containing the coding sequence for the gene of interest are transfected into cells, such as for example 293T cells, and supernatants collected after 48 hours. For cell treatment and RNA isolation, multiple primary human cells or human cell lines are used; such cells may include but are not limited to, Normal Human Dermal Fibroblasts, Aortic Smooth Muscle, Human Umbilical Vein Endothelial Cells, HepG2, Daudi, Jurkat, U937, Caco, and THP-1 cell lines. Cells are plated in growth media and growth is arrested by culturing without media change for 3 days, or by switching cells to low serum media and incubating overnight. Cells are treated for 1, 6, or 24 hours with either vector control supernatant or sample supernatant (or purified/partially purified protein preparations in buffer). Total RNA is isolated; for example, by using Trizol extraction or by using the Ambion RNAqueous™-4PCR RNA isolation system. Expression levels of multiple genes are analyzed using Taqman®, and expression in the test sample is compared to control vector samples to identify genes induced or repressed. Each of the above described techniques are well known to, and routinely performed by, those of ordinary skill in the art.

Table 1F indicates particular disease classes and preferred indications for which polynucleotides, polypeptides, or antibodies of the present invention may be used in detecting, diagnosing, preventing, treating and/or ameliorating said diseases and disorders based on “target” gene expression patterns which may be up- or down-regulated by polynucleotides (and the encoded polypeptides) corresponding to each indicated cDNA Clone ID (shown in Table 1F, Column 2).

Thus, in preferred embodiments, the present invention encompasses a method of detecting, diagnosing, preventing, treating, and/or ameliorating a disease or disorder listed in the “Disease Class” and/or “Preferred Indication” columns of Table 1F; comprising administering to a patient in which such detection, diagnosis, prevention, or treatment is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to detect, diagnose, prevent, treat, or ameliorate the disease or disorder. The first and second columns of Table 1D show the “Gene No.” and “cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may be used in detecting, diagnosing, preventing, treating, or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in the “Disease Class” or “Preferred Indication” Columns of Table 1F.

In another embodiment, the present invention also encompasses methods of detecting, diagnosing, preventing, treating, or ameliorating a disease or disorder listed in the “Disease Class” or “Preferred Indication” Columns of Table 1F; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in the “Disease Class” or “Preferred Indication” Columns of Table 1F.

The “Disease Class” Column of Table 1F provides a categorized descriptive heading for diseases, disorders, and/or conditions (more fully described below) that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The “Preferred Indication” Column of Table 1F describes diseases, disorders, and/or conditions that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The “Cell Line” and “Exemplary Targets” Columns of Table 1F indicate particular cell lines and target genes, respectively, which may show altered gene expression patterns (i.e., up- or down-regulation of the indicated target gene) in Taqman® assays, performed as described above, utilizing polynucleotides of the cDNA Clone ID shown in the corresponding row. Alteration of expression patterns of the indicated “Exemplary Target” genes is correlated with a particular “Disease Class” and/or “Preferred Indication” as shown in the corresponding row under the respective column headings.

The “Exemplary Accessions” Column indicates GenBank Accessions (available online through the National Center for Biotechnology Information (NCBI) at http://www.ncbi.nim.nih.gov/) which correspond to the “Exemplary Targets” shown in the adjacent row.

The recitation of “Cancer” in the “Disease Class” Column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate neoplastic diseases and/or disorders (e.g., leukemias, cancers, etc., as described below under “Hyperproliferative Disorders”).

The recitation of “Immune” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), blood disorders (e.g., as described below under “Immune Activity” “Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under “Infectious Disease”).

The recitation of “Angiogenesis” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), diseases and/or disorders of the cardiovascular system (e.g., as described below under “Cardiovascular Disorders”), diseases and/or disorders involving cellular and genetic abnormalities (e.g., as described below under “Diseases at the Cellular Level”), diseases and/or disorders involving angiogenesis (e.g., as described below under “Anti-Angiogenesis Activity”), to promote or inhibit cell or tissue regeneration (e.g., as described below under “Regeneration”), or to promote wound healing (e.g., as described below under “Wound Healing and Epithelial Cell Proliferation”).

The recitation of “Diabetes” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diabetes (including diabetes mellitus types I and II), as well as diseases and/or disorders associated with, or consequential to, diabetes (e.g. as described below under “Endocrine Disorders,” “Renal Disorders,” and “Gastrointestinal Disorders”).

Description of Table 1F

Polynucleotides encoding polypeptides of the present invention can be used in assays to test for one or more biological activities. One such biological activity which may be tested includes the ability of polynucleotides and polypeptides of the invention to stimulate up-regulation or down-regulation of expression of particular genes and proteins. Hence, if polynucleotides and polypeptides of the present invention exhibit activity in altering particular gene and protein expression patterns, it is likely that these polynucleotides and polypeptides of the present invention may be involved in, or capable of effecting changes in, diseases associated with the altered gene and protein expression profiles. Hence, polynucleotides, polypeptides, or antibodies of the present invention could be used to treat said associated diseases.

TaqMan® assays may be performed to assess the ability of polynucleotides (and polypeptides they encode) to alter the expression pattern of particular “target” genes. TaqMan® reactions are performed to evaluate the ability of a test agent to induce or repress expression of specific genes in different cell types. TaqMan® gene expression quantification assays (“TaqMan® assays”) are well known to, and routinely performed by, those of ordinary skill in the art. TaqMan® assays are performed in a two step reverse transcription/polymerase chain reaction (RT-PCR). In the first (RT) step, cDNA is reverse transcribed from total RNA samples using random hexamer primers. In the second (PCR) step, PCR products are synthesized from the cDNA using gene specific primers.

To quantify gene expression the Taqman® PCR reaction exploits the 5′ nuclease activity of AmpliTaq Gold® DNA Polymerase to cleave a Taqman® probe (distinct from the primers) during PCR. The Taqman® probe contains a reporter dye at the 5′-end of the probe and a quencher dye at the 3′ end of the probe. When the probe is intact, the proximity of the reporter dye to the quencher dye results in suppression of the reporter fluorescence. During PCR, if the target of interest is present, the probe specifically anneals between the forward and reverse primer sites. AmpliTaq Fold DNA Polymerase then cleaves the probe between the reporter and quencher when the probe hybridizes to the target, resulting in increased fluorescence of the reporter (see FIG. 2). Accumulation of PCR products is detected directly by monitoring the increase in fluorescence of the reporter dye.

After the probe fragments are displaced from the target, polymerization of the strand continues. The 3′-end of the probe is blocked to prevent extension of the probe during PCR. This process occurs in every cycle and does not interfere with the exponential accumulation of product. The increase in fluorescence signal is detected only if the target sequence is complementary to the probe and is amplified during PCR. Because of these requirements, any nonspecific amplification is not detected.

For test sample preparation, vector controls or constructs containing the coding sequence for the gene of interest are transfected into cells, such as for example 293T cells, and supernatants collected after 48 hours. For cell treatment and RNA isolation, multiple primary human cells or human cell lines are used; such cells may include but are not limited to, Normal Human Dermal Fibroblasts, Aortic Smooth Muscle, Human Umbilical Vein Endothelial Cells, HepG2, Daudi, Jurkat, U937, Caco, and THP-1 cell lines. Cells are plated in growth media and growth is arrested by culturing without media change for 3 days, or by switching cells to low serum media and incubating overnight. Cells are treated for 1, 6, or 24 hours with either vector control supernatant or sample supernatant (or purified/partially purified protein preparations in buffer). Total RNA is isolated; for example, by using Trizol extraction or by using the Ambion RNAqueous™-4PCR RNA isolation system. Expression levels of multiple genes are analyzed using Taqman®, and expression in the test sample is compared to control vector samples to identify genes induced or repressed. Each of the above described techniques are well known to, and routinely performed by, those of ordinary skill in the art.

Table 1F indicates particular disease classes and preferred indications for which polynucleotides, polypeptides, or antibodies of the present invention may be used in detecting, diagnosing, preventing, treating and/or ameliorating said diseases and disorders based on “target” gene expression patterns which may be up- or down-regulated by polynucleotides (and the encoded polypeptides) corresponding to each indicated cDNA Clone ID (shown in Table 1F, Column 2).

Thus, in preferred embodiments, the present invention encompasses a method of detecting, diagnosing, preventing, treating, and/or ameliorating a disease or disorder listed in the “Disease Class” and/or “Preferred Indication” columns of Table 1F; comprising administering to a patient in which such detection, diagnosis, prevention, or treatment is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to detect, diagnose, prevent, treat, or ameliorate the disease or disorder. The first and second columns of Table 1D show the “Gene No.” and “cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may be used in detecting, diagnosing, preventing, treating, or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in the “Disease Class” or “Preferred Indication” Columns of Table 1F.

In another embodiment, the present invention also encompasses methods of detecting, diagnosing, preventing, treating, or ameliorating a disease or disorder listed in the “Disease Class” or “Preferred Indication” Columns of Table 1F; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in the “Disease Class” or “Preferred Indication” Columns of Table 1F.

The “Disease Class” Column of Table 1F provides a categorized descriptive heading for diseases, disorders, and/or conditions (more fully described below) that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The “Preferred Indication” Column of Table 1F describes diseases, disorders, and/or conditions that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The “Cell Line” and “Exemplary Targets” Columns of Table 1F indicate particular cell lines and target genes, respectively, which may show altered gene expression patterns (i.e., up- or down-regulation of the indicated target gene) in Taqman® assays, performed as described above, utilizing polynucleotides of the cDNA Clone ID shown in the corresponding row. Alteration of expression patterns of the indicated “Exemplary Target” genes is correlated with a particular “Disease Class” and/or “Preferred Indication” as shown in the corresponding row under the respective column headings.

The “Exemplary Accessions” Column indicates GenBank Accessions (available online through the National Center for Biotechnology Information (NCBI) at http://www.ncbi.nlm.nih.gov/) which correspond to the “Exemplary Targets” shown in the adjacent row.

The recitation of “Cancer” in the “Disease Class” Column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate neoplastic diseases and/or disorders (e.g., leukemias, cancers, etc., as described below under “Hyperproliferative Disorders”).

The recitation of “Immune” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), blood disorders (e.g., as described below under “Immune Activity” “Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under “Infectious Disease”).

The recitation of “Angiogenesis” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), diseases and/or disorders of the cardiovascular system (e.g., as described below under “Cardiovascular Disorders”), diseases and/or disorders involving cellular and genetic abnormalities (e.g., as described below under “Diseases at the Cellular Level”), diseases and/or disorders involving angiogenesis (e.g., as described below under “Anti-Angiogenesis Activity”), to promote or inhibit cell or tissue regeneration (e.g., as described below under “Regeneration”), or to promote wound healing (e.g., as described below under “Wound Healing and Epithelial Cell Proliferation”).

The recitation of “Diabetes” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diabetes (including diabetes mellitus types I and II), as well as diseases and/or disorders associated with, or consequential to, diabetes (e.g. as described below under “Endocrine Disorders,” “Renal Disorders,” and “Gastrointestinal Disorders”).

Description of Table 2

Table 2 summarizes homology and features of some of the polypeptides of the invention. The first column provides a unique clone identifier, “Clone ID:”, corresponding to a cDNA clone disclosed in Table 1A or 1B. The second column provides the unique contig identifier, “Contig ID:” corresponding to contigs in Table 1B and allowing for correlation with the information in Table 1B. The third column provides the sequence identifier, “SEQ ID NO:X”, for the contig polynucleotide sequence. The fourth column provides the analysis method by which the homology/identity disclosed in the Table was determined. Comparisons were made between polypeptides encoded by the polynucleotides of the invention and either a non-redundant protein database (herein referred to as “NR”), or a database of protein families (herein referred to as “PFAM”) as further described below. The fifth column provides a description of the PFAM/NR hit having a significant match to a polypeptide of the invention. Column six provides the accession number of the PFAM/NR hit disclosed in the fifth column. Column seven, “Score/Percent Identity”, provides a quality score or the percent identity, of the hit disclosed in columns five and six. Columns 8 and 9, “NT From” and “NT To” respectively, delineate the polynucleotides in “SEQ ID NO:X” that encode a polypeptide having a significant match to the PFAM/NR database as disclosed in the fifth and sixth columns. In specific embodiments polypeptides of the invention comprise, or alternatively consist of, an amino acid sequence encoded by a polynucleotide in SEQ ID NO:X as delineated in columns 8 and 9, or fragments or variants thereof.

Description of Table 3

Table 3 provides polynucleotide sequences that may be disclaimed according to certain embodiments of the invention. The first column provides a unique clone identifier, “Clone ID”, for a cDNA clone related to contig sequences disclosed in Table 1B. The second column provides the sequence identifier, “SEQ ID NO:X”, for contig sequences disclosed in Table 1A and/or 1B. The third column provides the unique contig identifier, “Contig ID:”, for contigs disclosed in Table 1B. The fourth column provides a unique integer ‘a’ where ‘a’ is any integer between 1 and the final nucleotide minus 15 of SEQ ID NO:X, and the fifth column provides a unique integer ‘b’ where ‘b’ is any integer between 15 and the final nucleotide of SEQ ID NO:X, where both a and b correspond to the positions of nucleotide residues shown in SEQ ID NO:X, and where b is greater than or equal to a+14. For each of the polynucleotides shown as SEQ ID NO:X, the uniquely defined integers can be substituted into the general formula of a-b, and used to describe polynucleotides which may be preferably excluded from the invention. In certain embodiments, preferably excluded from the invention are at least one, two, three, four, five, ten, or more of the polynucleotide sequence(s) having the accession number(s) disclosed in the sixth column of this Table (including for example, published sequence in connection with a particular BAC clone). In further embodiments, preferably excluded from the invention are the specific polynucleotide sequence(s) contained in the clones corresponding to at least one, two, three, four, five, ten, or more of the available material having the accession numbers identified in the sixth column of this Table (including for example, the actual sequence contained in an identified BAC clone).

Description of Table 4

Table 4 provides a key to the tissue/cell source identifier code disclosed in Table 1B.2, column 5. Column 1 provides the tissue/cell source identifier code disclosed in Table 1B.2, Column 5. Columns 2-5 provide a description of the tissue or cell source. Note that “Description” and “Tissue” sources (i.e. columns 2 and 3) having the prefix “a_” indicates organs, tissues, or cells derived from “adult” sources. Codes corresponding to diseased tissues are indicated in column 6 with the word “disease.” The use of the word “disease” in column 6 is non-limiting. The tissue or cell source may be specific (e.g. a neoplasm), or may be disease-associated (e.g., a tissue sample from a normal portion of a diseased organ). Furthermore, tissues and/or cells lacking the “disease” designation may still be derived from sources directly or indirectly involved in a disease state or disorder, and therefore may have a further utility in that disease state or disorder. In numerous cases where the tissue/cell source is a library, column 7 identifies the vector used to generate the library.

Description of Table 5

Table 5 provides a key to the OMIM reference identification numbers disclosed in Table 1B.1, column 9. OMIM reference identification numbers (Table 5, Column 1) were derived from Online Mendelian Inheritance in Man (Online Mendelian Inheritance in Man, OMIM. McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University (Baltimore, Md.) and National Center for Biotechnology Information, National Library of Medicine, (Bethesda, Md.) 2000. World Wide Web URL: http://www.ncbi.nlm.nih.gov/omim/). Column 2 provides diseases associated with the cytologic band disclosed in Table 1B.1, column 8, as determined using the Morbid Map database.

Description of Table 6

Table 6 summarizes some of the ATCC Deposits, Deposit dates, and ATCC designation numbers of deposits made with the ATCC in connection with the present application. These deposits were made in addition to those described in the Table 1A.

Description of Table 7

Table 7 shows the cDNA libraries sequenced, and ATCC designation numbers and vector information relating to these cDNA libraries.

The first column shows the first four letters indicating the Library from which each library clone was derived. The second column indicates the catalogued tissue description for the corresponding libraries. The third column indicates the vector containing the corresponding clones. The fourth column shows the ATCC deposit designation for each library clone as indicated by the deposit information in Table 6.

Definitions

The following definitions are provided to facilitate understanding of certain terms used throughout this specification.

In the present invention, “isolated” refers to material removed from its original environment (e.g., the natural environment if it is naturally occurring), and thus is altered “by the hand of man” from its natural state. For example, an isolated polynucleotide could be part of a vector or a composition of matter, or could be contained within a cell, and still be “isolated” because that vector, composition of matter, or particular cell is not the original environment of the polynucleotide. The term “isolated” does not refer to genomic or cDNA libraries, whole cell total or mRNA preparations, genomic DNA preparations (including those separated by electrophoresis and transferred onto blots), sheared whole cell genomic DNA preparations or other compositions where the art demonstrates no distinguishing features of the polynucleotide/sequences of the present invention.

In the present invention, a “secreted” protein refers to those proteins capable of being directed to the ER, secretory vesicles, or the extracellular space as a result of a signal sequence, as well as those proteins released into the extracellular space without necessarily containing a signal sequence. If the secreted protein is released into the extracellular space, the secreted protein can undergo extracellular processing to produce a “mature” protein. Release into the extracellular space can occur by many mechanisms, including exocytosis and proteolytic cleavage.

As used herein, a “polynucleotide” refers to a molecule having a nucleic acid sequence encoding SEQ ID NO:Y or a fragment or variant thereof (e.g., the polypeptide delinated in columns fourteen and fifteen of Table 1A); a nucleic acid sequence contained in SEQ ID NO:X (as described in column 5 of Table 1A and/or column 3 of Table 1B) or the complement thereof; a cDNA sequence contained in Clone ID: (as described in column 2 of Table 1A and/or 1B and contained within a library deposited with the ATCC); a nucleotide sequence encoding the polypeptide encoded by a nucleotide sequence in SEQ ID NO:B as defined in column 6 (EXON From-To) of Table 1C or a fragment or variant thereof; or a nucleotide coding sequence in SEQ ID NO:B as defined in column 6 of Table 1C or the complement thereof. For example, the polynucleotide can contain the nucleotide sequence of the full length cDNA sequence, including the 5′ and 3′ untranslated sequences, the coding region, as well as fragments, epitopes, domains, and variants of the nucleic acid sequence. Moreover, as used herein, a “polypeptide” refers to a molecule having an amino acid sequence encoded by a polynucleotide of the invention as broadly defined (obviously excluding poly-Phenylalanine or poly-Lysine peptide sequences which result from translation of a polyA tail of a sequence corresponding to a cDNA).

In the present invention, “SEQ ID NO:X” was often generated by overlapping sequences contained in multiple clones (contig analysis). A representative clone containing all or most of the sequence for SEQ ID NO:X is deposited at Human Genome Sciences, Inc. (HGS) in a catalogued and archived library. As shown, for example, in column 2 of Table 1B, each clone is identified by a cDNA Clone ID (identifier generally referred to herein as Clone ID:). Each Clone ID is unique to an individual clone and the Clone ID is all the information needed to retrieve a given clone from the HGS library. Table 7 provides a list of the deposited cDNA libraries. One can use the Clone ID: to determine the library source by reference to Tables 6 and 7. Table 7 lists the deposited cDNA libraries by name and links each library to an ATCC Deposit. Library names contain four characters, for example, “HTWE.” The name of a cDNA clone (Clone ID) isolated from that library begins with the same four characters, for example “HTWEP07”. As mentioned below, Table 1A and/or 1B correlates the Clone ID names with SEQ ID NO:X. Thus, starting with an SEQ ID NO:X, one can use Tables 1A, 1B, 6, 7, and 9 to determine the corresponding Clone ID, which library it came from and which ATCC deposit the library is contained in. Furthermore, it is possible to retrieve a given cDNA clone from the source library by techniques known in the art and described elsewhere herein. The ATCC is located at 10801 University Boulevard, Manassas, Va. 20110-2209, USA. The ATCC deposits were made pursuant to the terms of the Budapest Treaty on the international recognition of the deposit of microorganisms for the purposes of patent procedure.

In specific embodiments, the polynucleotides of the invention are at least 15, at least 30, at least 50, at least 100, at least 125, at least 500, or at least 1000 continuous nucleotides but are less than or equal to 300 kb, 200 kb, 100 kb, 50 kb, 15 kb, 10 kb, 7.5 kb, 5 kb, 2.5 kb, 2.0 kb, or 1 kb, in length. In a further embodiment, polynucleotides of the invention comprise a portion of the coding sequences, as disclosed herein, but do not comprise all or a portion of any intron. In another embodiment, the polynucleotides comprising coding sequences do not contain coding sequences of a genomic flanking gene (i.e., 5′ or 3′ to the gene of interest in the genome). In other embodiments, the polynucleotides of the invention do not contain the coding sequence of more than 1000, 500, 250, 100, 50, 25, 20, 15, 10, 5, 4, 3, 2, or 1 genomic flanking gene(s).

A “polynucleotide” of the present invention also includes those polynucleotides capable of hybridizing, under stringent hybridization conditions, to sequences contained in SEQ ID NO:X, or the complement thereof (e.g., the complement of any one, two, three, four, or more of the polynucleotide fragments described herein), the polynucleotide sequence delineated in columns 7 and 8 of Table 1A or the complement thereof, the polynucleotide sequence delineated in columns 8 and 9 of Table 2 or the complement thereof, and/or cDNA sequences contained in Clone ID: (e.g., the complement of any one, two, three, four, or more of the polynucleotide fragments, or the cDNA clone within the pool of cDNA clones deposited with the ATCC, described herein), and/or the polynucleotide sequence delineated in column 6 of Table 1C or the complement thereof. “Stringent hybridization conditions” refers to an overnight incubation at 42 degree C. in a solution comprising 50% formamide, 5×SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5× Denhardt's solution, 10% dextran sulfate, and 20 μg/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at about 65 degree C.

Also contemplated are nucleic acid molecules that hybridize to the polynucleotides of the present invention at lower stringency hybridization conditions. Changes in the stringency of hybridization and signal detection are primarily accomplished through the manipulation of formamide concentration (lower percentages of formamide result in lowered stringency); salt conditions, or temperature. For example, lower stringency conditions include an overnight incubation at 37 degree C. in a solution comprising 6×SSPE (20×SSPE=3M NaCl; 0.2M NaH₂PO₄; 0.02M EDTA, pH 7.4), 0.5% SDS, 30% formamide, 100 ug/ml salmon sperm blocking DNA; followed by washes at 50 degree C. with 1×SSPE, 0.1% SDS. In addition, to achieve even lower stringency, washes performed following stringent hybridization can be done at higher salt concentrations (e.g. 5×SSC).

Note that variations in the above conditions may be accomplished through the inclusion and/or substitution of alternate blocking reagents used to suppress background in hybridization experiments. Typical blocking reagents include Denhardt's reagent, BLOTTO, heparin, denatured salmon sperm DNA, and commercially available proprietary formulations. The inclusion of specific blocking reagents may require modification of the hybridization conditions described above, due to problems with compatibility.

Of course, a polynucleotide which hybridizes only to polyA+ sequences (such as any 3′ terminal polyA+ tract of a cDNA shown in the sequence listing), or to a complementary stretch of T (or U) residues, would not be included in the definition of “polynucleotide,” since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (e.g., practically any double-stranded cDNA clone generated using oligo dT as a primer).

The polynucleotide of the present invention can be composed of any polyribonucleotide or polydeoxribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. For example, polynucleotides can be composed of single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. In addition, the polynucleotide can be composed of triple-stranded regions comprising RNA or DNA or both RNA and DNA. A polynucleotide may also contain one or more modified bases or DNA or RNA backbones modified for stability or for other reasons. “Modified” bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, “polynucleotide” embraces chemically, enzymatically, or metabolically modified forms.

In specific embodiments, the polynucleotides of the invention are at least 15, at least 30, at least 50, at least 100, at least 125, at least 500, or at least 1000 continuous nucleotides but are less than or equal to 300 kb, 200 kb, 100 kb, 50 kb, 15 kb, 10 kb, 7.5 kb, 5 kb, 2.5 kb, 2.0 kb, or 1 kb, in length. In a further embodiment, polynucleotides of the invention comprise a portion of the coding sequences, as disclosed herein, but do not comprise all or a portion of any intron. In another embodiment, the polynucleotides comprising coding sequences do not contain coding sequences of a genomic flanking gene (i.e., 5′ or 3′ to the gene of interest in the genome). In other embodiments, the polynucleotides of the invention do not contain the coding sequence of more than 1000, 500, 250, 100, 50, 25, 20, 15, 10, 5, 4, 3, 2, or 1 genomic flanking gene(s).

“SEQ ID NO:X” refers to a polynucleotide sequence described in column 5 of Table 1A, while “SEQ ID NO:Y” refers to a polypeptide sequence described in column 10 of Table 1A. SEQ ID NO:X is identified by an integer specified in column 6 of Table 1A. The polypeptide sequence SEQ ID NO:Y is a translated open reading frame (ORF) encoded by polynucleotide SEQ ID NO:X. The polynucleotide sequences are shown in the sequence listing immediately followed by all of the polypeptide sequences. Thus, a polypeptide sequence corresponding to polynucleotide sequence SEQ ID NO:2 is the first polypeptide sequence shown in the sequence listing. The second polypeptide sequence corresponds to the polynucleotide sequence shown as SEQ ID NO:3, and so on.

The polypeptide of the present invention can be composed of amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain amino acids other than the 20 gene-encoded amino acids. The polypeptides may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications. Polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. (See, for instance, PROTEINS—STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993); POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, pgs. 1-12 (1983); Seifter et al., Meth. Enzymol. 182:626-646 (1990); Rattan et al., Ann. N.Y. Acad. Sci. 663:48-62 (1992)).

“SEQ ID NO:X” refers to a polynucleotide sequence described, for example, in Tables 1A, 1B or 2, while “SEQ ID NO:Y” refers to a polypeptide sequence described in column II of Table 1A and or column 6 of Table 1B.1. SEQ ID NO:X is identified by an integer specified in column 4 of Table 1B.1. The polypeptide sequence SEQ ID NO:Y is a translated open reading frame (ORF) encoded by polynucleotide SEQ ID NO:X. “Clone ID:” refers to a cDNA clone described in column 2 of Table 1A and/or 1B.

“A polypeptide having functional activity” refers to a polypeptide capable of displaying one or more known functional activities associated with a full-length (complete) protein. Such functional activities include, but are not limited to, biological activity, antigenicity [ability to bind (or compete with a polypeptide for binding) to an anti-polypeptide antibody], immunogenicity (ability to generate antibody which binds to a specific polypeptide of the invention), ability to form multimers with polypeptides of the invention, and ability to bind to a receptor or ligand for a polypeptide.

The polypeptides of the invention can be assayed for functional activity (e.g. biological activity) using or routinely modifying assays known in the art, as well as assays described herein. Specifically, one of skill in the art may routinely assay secreted polypeptides (including fragments and variants) of the invention for activity using assays as described in the examples section below.

“A polypeptide having biological activity” refers to a polypeptide exhibiting activity similar to, but not necessarily identical to, an activity of a polypeptide of the present invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. In the case where dose dependency does exist, it need not be identical to that of the polypeptide, but rather substantially similar to the dose-dependence in a given activity as compared to the polypeptide of the present invention (i.e., the candidate polypeptide will exhibit greater activity or not more than about 25-fold less and, preferably, not more than about tenfold less activity, and most preferably, not more than about three-fold less activity relative to the polypeptide of the present invention).

Tables

Table 1A

Table 1A summarizes information concerning certain polynucleotides and polypeptides of the invention. The first column provides the gene number in the application for each clone identifier. The second column provides a unique clone identifier, “Clone ID:”, for a cDNA clone related to each contig sequence disclosed in Table 1A. Third column, the cDNA Clones identified in the second column were deposited as indicated in the third column (i.e. by ATCC Deposit No:Z and deposit date). Some of the deposits contain multiple different clones corresponding to the same gene. In the fourth column, “Vector” refers to the type of vector contained in the corresponding cDNA Clone identified in the second column. In the fifth column, the nucleotide sequence identified as “NT SEQ ID NO:X” was assembled from partially homologous (“overlapping”) sequences obtained from the corresponding cDNA clone identified in the second column and, in some cases, from additional related cDNA clones. The overlapping sequences were assembled into a single contiguous sequence of high redundancy (usually three to five overlapping sequences at each nucleotide position), resulting in a final sequence identified as SEQ ID NO:X. In the sixth column, “Total NT Seq.” refers to the total number of nucleotides in the contig sequence identified as SEQ ID NO:X.” The deposited clone may contain all or most of these sequences, reflected by the nucleotide position indicated as “5′ NT of Clone Seq.” (seventh column) and the “3′ NT of Clone Seq.” (eighth column) of SEQ ID NO:X. In the ninth column, the nucleotide position of SEQ ID NO:X of the putative start codon (methionine) is identified as “5′ NT of Start Codon.” Similarly, in column ten, the nucleotide position of SEQ ID NO:X of the predicted signal sequence is identified as “5′ NT of First AA of Signal Pep.” In the eleventh column, the translated amino acid sequence, beginning with the methionine, is identified as “AA SEQ ID NO:Y,” although other reading frames can also be routinely translated using known molecular biology techniques. The polypeptides produced by these alternative open reading frames are specifically contemplated by the present invention.

In the twelfth and thirteenth columns of Table 1A, the first and last amino acid position of SEQ ID NO:Y of the predicted signal peptide is identified as “First AA of Sig Pep” and “Last AA of Sig Pep.” In the fourteenth column, the predicted first amino acid position of SEQ ID NO:Y of the secreted portion is identified as “Predicted First AA of Secreted Portion”. The amino acid position of SEQ ID NO:Y of the last amino acid encoded by the open reading frame is identified in the fifteenth column as “Last AA of ORF”.

SEQ ID NO:X (where X may be any of the polynucleotide sequences disclosed in the sequence listing) and the translated SEQ ID NO:Y (where Y may be any of the polypeptide sequences disclosed in the sequence listing) are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below. For instance, SEQ ID NO:X is useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ID NO:X or the cDNA contained in the deposited clone. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling a variety of forensic and diagnostic methods of the invention. Similarly, polypeptides identified from SEQ ID NO:Y may be used, for example, to generate antibodies which bind specifically to proteins containing the polypeptides and the secreted proteins encoded by the cDNA clones identified in Table 1A and/or elsewhere herein

Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence. In these cases, the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).

Accordingly, for those applications requiring precision in the nucleotide sequence or the amino acid sequence, the present invention provides not only the generated nucleotide sequence identified as SEQ ID NO:X, and the predicted translated amino acid sequence identified as SEQ ID NO:Y, but also a sample of plasmid DNA containing a human cDNA of the invention deposited with the ATCC, as set forth in Table 1A. The nucleotide sequence of each deposited plasmid can readily be determined by sequencing the deposited plasmid in accordance with known methods

The predicted amino acid sequence can then be verified from such deposits. Moreover, the amino acid sequence of the protein encoded by a particular plasmid can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human cDNA, collecting the protein, and determining its sequence.

Also provided in Table 1A is the name of the vector which contains the cDNA plasmid. Each vector is routinely used in the art. The following additional information is provided for convenience.

Vectors Lambda Zap (U.S. Pat. Nos. 5,128,256 and 5,286,636), Uni-Zap XR (U.S. Pat. Nos. 5,128,256 and 5,286,636), Zap Express (U.S. Pat. Nos. 5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. et al., Nucleic Acids Res. 16:7583-7600 (1988); Alting-Mees, M. A. and Short, J. M., Nucleic Acids Res. 17:9494 (1989)) and pBK (Alting-Mees, M. A. et al., Strategies 5:58-61 (1992)) are commercially available from Stratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La Jolla, Calif., 92037. pBS contains an ampicillin resistance gene and pBK contains a neomycin resistance gene. Phagemid pBS may be excised from the Lambda Zap and Uni-Zap XR vectors, and phagemid pBK may be excised from the Zap Express vector. Both phagemids may be transformed into E. coli strain XL-1 Blue, also available from Stratagene

Vectors pSport1, pCMVSport 1.0, pCMVSport 2.0 and pCMVSport 3.0, were obtained from Life Technologies, Inc., P.O. Box 6009, Gaithersburg, Md. 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. See, for instance, Gruber, C. E., et al., Focus 15:59 (1993). Vector lafmid BA (Bento Soares, Columbia University, New York, N.Y.) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue. Vector pCR®2.1, which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, Calif. 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. See, for instance, Clark, J. M., Nuc. Acids Res. 16:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991).

The present invention also relates to the genes corresponding to SEQ ID NO:X, SEQ ID NO:Y, and/or a deposited cDNA (cDNA Clone ID). The corresponding gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include, but are not limited to, preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material.

Also provided in the present invention are allelic variants, orthologs, and/or species homologs. Procedures known in the art can be used to obtain full-length genes, allelic variants, splice variants, full-length coding portions, orthologs, and/or species homologs of genes corresponding to SEQ ID NO:X and SEQ ID NO:Y using information from the sequences disclosed herein or the clones deposited with the ATCC. For example, allelic variants and/or species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for allelic variants and/or the desired homologue.

The present invention provides a polynucleotide comprising, or alternatively consisting of, the nucleic acid sequence of SEQ ID NO:X and/or a cDNA contained in ATCC Deposit No.Z. The present invention also provides a polypeptide comprising, or alternatively, consisting of, the polypeptide sequence of SEQ ID NO:Y, a polypeptide encoded by SEQ ID NO:X, and/or a polypeptide encoded by a cDNA contained in ATCC deposit No.Z. Polynucleotides encoding a polypeptide comprising, or alternatively consisting of the polypeptide sequence of SEQ ID NO:Y, a polypeptide encoded by SEQ ID NO:X and/or a polypeptide encoded by the cDNA contained in ATCC Deposit No.Z, are also encompassed by the invention. The present invention further encompasses a polynucleotide comprising, or alternatively consisting of the complement of the nucleic acid sequence of SEQ ID NO:X, and/or the complement of the coding strand of the cDNA contained in ATCC Deposit No.Z. LENGTHY TABLE REFERENCED HERE US20070032413A1-20070208-T00001 Please refer to the end of the specification for access instructions. Table 1B (Comprised of Tables 1B.1 and 1B.2)

The first column in Table 1B.1 and Table 1B.2 provides the gene number in the application corresponding to the clone identifier. The second column in Table 1B.1 and Table 1B.2 provides a unique “Clone ID:” for the cDNA clone related to each contig sequence disclosed in Table 1B.1 and Table 1B.2. This clone ID references the cDNA clone which contains at least the 5′ most sequence of the assembled contig and at least a portion of SEQ ID NO:X as determined by directly sequencing the referenced clone. The referenced clone may have more sequence than described in the sequence listing or the clone may have less. In the vast majority of cases, however, the clone is believed to encode a full-length polypeptide. In the case where a clone is not full-length, a full-length cDNA can be obtained by methods described elsewhere herein. The third column in Table 1B.1 and Table 1B.2 provides a unique “Contig ID” identification for each contig sequence. The fourth column in Table 1B.1 and Table 1B.2 provides the “SEQ ID NO:” identifier for each of the contig polynucleotide sequences disclosed in Table 1B.

Table 1B.1

The fifth column in Table 1B.1, “ORF (From-To)”, provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence “SEQ ID NO:X” that delineate the preferred open reading frame (ORF) shown in the sequence listing and referenced in Table 1B.1, column 6, as SEQ ID NO:Y. Where the nucleotide position number “To” is lower than the nucleotide position number “From”, the preferred ORF is the reverse complement of the referenced polynucleotide sequence. The sixth column in Table 1B.1 provides the corresponding SEQ ID NO:Y for the polypeptide sequence encoded by the preferred ORF delineated in column 5. In one embodiment, the invention provides an amino acid sequence comprising, or alternatively consisting of, a polypeptide encoded by the portion of SEQ ID NO:X delineated by “ORF (From-To)”. Also provided are polynucleotides encoding such amino acid sequences and the complementary strand thereto. Column 7 in Table 1B.1 lists residues comprising epitopes contained in the polypeptides encoded by the preferred ORF (SEQ ID NO:Y), as predicted using the algorithm of Jameson and Wolf, (1988) Comp. Appl. Biosci. 4:181-186. The Jameson-Wolf antigenic analysis was performed using the computer program PROTEAN (Version 3.11 for the Power MacIntosh, DNASTAR, Inc., 1228 South Park Street Madison, Wis.). In specific embodiments, polypeptides of the invention comprise, or alternatively consist of, at least one, two, three, four, five or more of the predicted epitopes as described in Table 1B. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly.

Column 8 in Table 1B.1 provides a chromosomal map location for certain polynucleotides of the invention. Chromosomal location was determined by finding exact matches to EST and cDNA sequences contained in the NCBI (National Center for Biotechnology Information) UniGene database. Each sequence in the UniGene database is assigned to a “cluster”; all of the ESTs, cDNAs, and STSs in a cluster are believed to be derived from a single gene. Chromosomal mapping data is often available for one or more sequence(s) in a UniGene cluster; this data (if consistent) is then applied to the cluster as a whole. Thus, it is possible to infer the chromosomal location of a new polynucleotide sequence by determining its identity with a mapped UniGene cluster.

A modified version of the computer program BLASTN (Altshul, et al., J. Mol. Biol. 215:403-410 (1990), and Gish, and States, Nat. Genet. 3:266-272) (1993) was used to search the UniGene database for EST or cDNA sequences that contain exact or near-exact matches to a polynucleotide sequence of the invention (the ‘Query’). A sequence from the UniGene database (the ‘Subject’) was said to be an exact match if it contained a segment of 50 nucleotides in length such that 48 of those nucleotides were in the same order as found in the Query sequence. If all of the matches that met this criteria were in the same UniGene cluster, and mapping data was available for this cluster, it is indicated in Table 1B under the heading “Cytologic Band”. Where a cluster had been further localized to a distinct cytologic band, that band is disclosed; where no banding information was available, but the gene had been localized to a single chromosome, the chromosome is disclosed.

Once a presumptive chromosomal location was determined for a polynucleotide of the invention, an associated disease locus was identified by comparison with a database of diseases which have been experimentally associated with genetic loci. The database used was the Morbid Map, derived from OMIM™ and National Center for Biotechnology Information, National Library of Medicine (Bethesda, Md.) 2000. If the putative chromosomal location of a polynucleotide of the invention (Query sequence) was associated with a disease in the Morbid Map database, an OMIM reference identification number was noted in column 9, Table 1B.1, labelled “OMIM Disease Reference(s). Table 5 is a key to the OMIM reference identification numbers (column 1), and provides a description of the associated disease in Column 2.

Table 1B.2

Column 5, in Table 1B.2, provides an expression profile and library Code: Count for each of the contig sequences (SEQ ID NO:X) disclosed in Table 1B, which can routinely be combined with the information provided in Table 4 and used to determine the tissues, cells, and/or cell linc libraries which predominantly express the polynucleotides of the invention. The first number in Table 1B.2, column 5 (preceding the colon), represents the tissue/cell source identifier code corresponding to the code and description provided in Table 4. The second number in column 5 (following the colon) represents the number of times a sequence corresponding to the reference polynucleotide sequence was identified in the corresponding tissue/cell source. Those tissue/cell source identifier codes in which the first two letters are “AR” designate information generated using DNA array technology. Utilizing this technology, cDNAs were amplified by PCR and then transferred, in duplicate, onto the array. Gene expression was assayed through hybridization of first strand cDNA probes to the DNA array. cDNA probes were generated from total RNA extracted from a variety of different tissues and cell lines. Probe synthesis was performed in the presence of ³³P dCTP, using oligo (dT) to prime reverse transcription. After hybridization, high stringency washing conditions were employed to remove non-specific hybrids from the array. The remaining signal, emanating from each gene target, was measured using a Phosphorimager. Gene expression was reported as Phosphor Stimulating Luminescence (PSL) which reflects the level of phosphor signal generated from the probe hybridized to each of the gene targets represented on the array. A local background signal subtraction was performed before the total signal generated from each array was used to normalize gene expression between the different hybridizations. The value presented after “[array code]:” represents the mean of the duplicate values, following background subtraction and probe normalization. One of skill in the art could routinely use this information to identify normal and/or diseased tissue(s) which show a predominant expression pattern of the corresponding polynucleotide of the invention or to identify polynucleotides which show predominant and/or specific tissue and/or cell expression. LENGTHY TABLE REFERENCED HERE US20070032413A1-20070208-T00002 Please refer to the end of the specification for access instructions.

Tables 1D and 1E: The polynucleotides or polypeptides, or agonists or antagonists of the present invention can be used in assays to test for one or more biological activities. If these polynucleotides and polypeptides do exhibit activity in a particular assay, it is likely that these molecules may be involved in the diseases associated with the biological activity. Thus, the polynucleotides or polypeptides, or agonists or antagonists could be used to treat the associated disease.

The present invention encompasses methods of detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating a disease or disorder. In preferred embodiments, the present invention encompasses a method of treating a disease or disorder listed in the “Preferred Indications” columns of Table 1D and Table 1E; comprising administering to a patient (in which such treatment, prevention, or amelioration is desired) a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to treat, prevent, diagnose, or ameliorate the disease or disorder. The first and second columns of Table 1D show the “Gene No.” and “cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may be used in preventing, treating, diagnosing, or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in Column 3 of Table 1D.

In another embodiment, the present invention also encompasses methods of preventing, treating, diagnosing, or ameliorating a disease or disorder listed in the “Preferred Indications” column of Table 1D and Table 1E; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in Column 3 of Table 1D.

The “Preferred Indications” columns of Table 1D and Table 1E describe diseases, disorders, and/or conditions that may be treated, prevented, diagnosed, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The recitation of “Cancer” in the “Preferred Indications” columns indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., leukemias, cancers, and/or as described below under “Hyperproliferative Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D may be used for example, to diagnose, treat, prevent, and/or ameliorate a neoplasm located in a tissue selected from the group consisting of: colon, abdomen, bone, breast, digestive system, liver, pancreas, prostate, peritoneum, lung, blood (e.g., leukemia), endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid), uterus, eye, head and neck, nervous (central and peripheral), lymphatic system, pelvic, skin, soft tissue, spleen, thoracic, and urogenital.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a pre-neoplastic condition, selected from the group consisting of: hyperplasia (e.g., endometrial hyperplasia and/or as described in the section entitled “Hyperproliferative Disorders”), metaplasia (e.g., connective tissue metaplasia, atypical metaplasia, and/or as described in the section entitled “Hyperproliferative Disorders”), and/or dysplasia (e.g., cervical dysplasia, and bronchopulmonary dysplasia).

In another specific embodiment, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a benign dysproliferative disorder selected from the group consisting of: benign tumors, fibrocystic conditions, tissue hypertrophy, and/or as described in the section entitled “Hyperproliferative Disorders”.

The recitation of “Immune/Hematopoietic” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), blood disorders (e.g., as described below under “Immune Activity” “Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under “Infectious Disease”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having the “Immune/Hematopoietic” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: anemia, pancytopenia, leukopenia, thrombocytopenia, leukemias, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphocytic anemia (ALL), plasmacytomas, multiple myeloma, Burkitt's lymphoma, arthritis, asthma, AIDS, autoimmune disease, rheumatoid arthritis, granulomatous disease, immune deficiency, inflammatory bowel disease, sepsis, neutropenia, neutrophilia, psoriasis, immune reactions to transplanted organs and tissues, systemic lupus erythematosis, hemophilia, hypercoagulation, diabetes mellitus, endocarditis, meningitis, Lyme Disease, and allergies.

The recitation of “Reproductive” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the reproductive system (e.g., as described below under “Reproductive System Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Reproductive” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cryptorchism, prostatitis, inguinal hernia, varicocele, leydig cell tumors, verrucous carcinoma, prostatitis, malacoplakia, Peyronie's disease, penile carcinoma, squamous cell hyperplasia, dysmenorrhea, ovarian adenocarcinoma, Turner's syndrome, mucopurulent cervicitis, Sertoli-leydig tumors, ovarian cancer, uterine cancer, pelvic inflammatory disease, testicular cancer, prostate cancer, Klinefelter's syndrome, Young's syndrome, premature ejaculation, diabetes mellitus, cystic fibrosis, Kartagener's syndrome, testicular atrophy, testicular feminization, anorchia, ectopic testis, epididymitis, orchitis, gonorrhea, syphilis, testicular torsion, vasitis nodosa, germ cell tumors, stromal tumors, dysmenorrhea, retroverted uterus, endometriosis, fibroids, adenomyosis, anovulatory bleeding, amenorrhea, Cushing's syndrome, hydatidiform moles, Asherman's syndrome, premature menopause, precocious puberty, uterine polyps, dysfunctional uterine bleeding, cervicitis, chronic cervicitis, mucopurulent cervicitis, cervical dysplasia, cervical polyps, Nabothian cysts, cervical erosion, cervical incompetence, cervical neoplasms, pseudohermaphroditism, and premenstrual syndrome.

The recitation of “Musculoskeletal” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or valiant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the immune system (e.g., as described below under “Immune Activity”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Musculoskeletal” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: bone cancers (e.g., osteochondromas, benign chondromas, chondroblastoma, chondromyxoid fibromas, osteoid osteomas, giant cell tumors, multiple myeloma, osteosarcomas), Paget's Disease, rheumatoid arthritis, systemic lupus erythematosus, osteomyelitis, Lyme Disease, gout, bursitis, tendonitis, osteoporosis, osteoarthritis, muscular dystrophy, mitochondrial myopathy, cachexia, and multiple sclerosis.

The recitation of “Cardiovascular” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the cardiovascular system (e.g., as described below under “Cardiovascular Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cardiovascular” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: myxomas, fibromas, rhabdomyomas, cardiovascular abnormalities (e.g., congenital heart defects, cerebral arteriovenous malformations, septal defects), heart disease (e.g., heart failure, congestive heart disease, arrhythmia, tachycardia, fibrillation, pericardial Disease, endocarditis), cardiac arrest, heart valve disease (e.g., stenosis, regurgitation, prolapse), vascular disease (e.g., hypertension, coronary artery disease, angina, aneurysm, arteriosclerosis, peripheral vascular disease), hyponatremia, hypernatremia, hypokalemia, and hyperkalemia.

The recitation of “Mixed Fetal” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Mixed Fetal” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: spina bifida, hydranencephaly, neurofibromatosis, fetal alcohol syndrome, diabetes mellitus, PKU, Down's syndrome, Patau syndrome, Edwards syndrome, Turner syndrome, Apert syndrome, Carpenter syndrome, Conradi syndrome, Crouzon syndrome, cutis laxa, Cornelia de Lange syndrome, Ellis-van Creveld syndrome, Holt-Oram syndrome, Kartagener syndrome, Meckel-Gruber syndrome, Noonan syndrome, Pallister-Hall syndrome, Rubinstein-Taybi syndrome, Scimitar syndrome, Smith-Lemli-Opitz syndrome, thromocytopenia-absent radius (TAR) syndrome, Treacher Collins syndrome, Williams syndrome, Hirschsprung's disease, Meckel's diverticulum, polycystic kidney disease, Turner's syndrome, and gonadal dysgenesis, Klippel-Feil syndrome, Ostogenesis imperfecta, muscular dystrophy, Tay-Sachs disease, Wilm's tumor, neuroblastoma, and retinoblastoma.

The recitation of “Excretory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and renal disorders (e.g., as described below under “Renal Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Excretory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: bladder cancer, prostate cancer, benign prostatic hyperplasia, bladder disorders (e.g., urinary incontinence, urinary retention, urinary obstruction, urinary tract Infections, interstitial cystitis, prostatitis, neurogenic bladder, hematuria), renal disorders (e.g., hydronephrosis, proteinuria, renal failure, pyelonephritis, urolithiasis, reflux nephropathy, and unilateral obstructive uropathy).

The recitation of “Neural/Sensory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the nervous system (e.g., as described below under “Neural Activity and Neurological Diseases”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Neural/Sensory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: brain cancer (e.g., brain stem glioma, brain tumors, central nervous system (Primary) lymphoma, central nervous system lymphoma, cerebellar astrocytoma, and cerebral astrocytoma, neurodegenerative disorders (e.g., Alzheimer's Disease, Creutzfeldt-Jakob Disease, Parkinson's Disease, and Idiopathic Presenile Dementia), encephalomyelitis, cerebral malaria, meningitis, metabolic brain diseases (e.g., phenylketonuria and pyruvate carboxylase deficiency), cerebellar ataxia, ataxia telangiectasia, and AIDS Dementia Complex, schizophrenia, attention deficit disorder, hyperactive attention deficit disorder, autism, and obsessive compulsive disorders.

The recitation of “Respiratory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the respiratory system (e.g., as described below under “Respiratory Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Respiratory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cancers of the respiratory system such as larynx cancer, pharynx cancer, trachea cancer, epiglottis cancer, lung cancer, squamous cell carcinomas, small cell (oat cell) carcinomas, large cell carcinomas, and adenocarcinomas. Allergic reactions, cystic fibrosis, sarcoidosis, histiocytosis X, infiltrative lung diseases (e.g., pulmonary fibrosis and lymphoid interstitial pneumonia), obstructive airway diseases (e.g., asthma, emphysema, chronic or acute bronchitis), occupational lung diseases (e.g., silicosis and asbestosis), pneumonia, and pleurisy.

The recitation of “Endocrine” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the respiratory system (e.g., as described below under “Respiratory Disorders”), renal disorders (e.g., as described below under “Renal Disorders”), and disorders of the endocrine system (e.g., as described below under “Endocrine Disorders”.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having an “Endocrine” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cancers of endocrine tissues and organs (e.g., cancers of the hypothalamus, pituitary gland, thyroid gland, parathyroid glands, pancreas, adrenal glands, ovaries, and testes), diabetes (e.g., diabetes insipidus, type I and type II diabetes mellitus), obesity, disorders related to pituitary glands (e.g., hyperpituitarism, hypopituitarism, and pituitary dwarfism), hypothyroidism, hyperthyroidism, goiter, reproductive disorders (e.g. male and female infertility), disorders related to adrenal glands (e.g., Addison's Disease, corticosteroid deficiency, and Cushing's Syndrome), kidney cancer (e.g., hypemephroma, transitional cell cancer, and Wilm's tumor), diabetic nephropathy, interstitial nephritis, polycystic kidney disease, glomerulonephritis (e.g., IgM mesangial proliferative glomerulonephritis and glomerulonephritis caused by autoimmune disorders; such as Goodpasture's syndrome), and nephrocalcinosis.

The recitation of “Digestive” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the gastrointestinal system (e.g., as described below under “Gastrointestinal Disorders”.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Digestive” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: ulcerative colitis, appendicitis, Crohn's disease, hepatitis, hepatic encephalopathy, portal hypertension, cholelithiasis, cancer of the digestive system (e.g., biliary tract cancer, stomach cancer, colon cancer, gastric cancer, pancreatic cancer, cancer of the bile duct, tumors of the colon (e.g., polyps or cancers), and cirrhosis), pancreatitis, ulcerative disease, pyloric stenosis, gastroenteritis, gastritis, gastric atropy, benign tumors of the duodenum, distension, irritable bowel syndrome, malabsorption, congenital disorders of the small intestine, bacterial and parasitic infection, megacolon, Hirschsprung's disease, aganglionic megacolon, acquired megacolon, colitis, anorectal disorders (e.g., anal fistulas, hemorrhoids), congenital disorders of the liver (e.g., Wilson's disease, hemochromatosis, cystic fibrosis, biliary atresia, and alpha1-antitrypsin deficiency), portal hypertension, cholelithiasis, and jaundice.

The recitation of “Connective/Epithelial” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), cellular and genetic abnormalities (e.g., as described below under “Diseases at the Cellular Level”), angiogenesis (e.g., as described below under “Anti-Angiogenesis Activity”), and or to promote or inhibit regeneration (e.g., as described below under “Regeneration”), and wound healing (e.g., as described below under “Wound Healing and Epithelial Cell Proliferation”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Connective/Epithelial” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: connective tissue metaplasia, mixed connective tissue disease, focal epithelial hyperplasia, epithelial metaplasia, mucoepithelial dysplasia, graft v. host disease, polymyositis, cystic hyperplasia, cerebral dysplasia, tissue hypertrophy, Alzheimer's disease, lymphoproliferative disorder, Waldenstron's macroglobulinemia, Crohn's disease, pernicious anemia, idiopathic Addison's disease, glomerulonephritis, bullous pemphigoid, Sjogren's syndrome, diabetes mellitus, cystic fibrosis, osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, osteoporosis, osteocarthritis, periodontal disease, wound healing, relapsing polychondritis, vasculitis, polyarteritis nodosa, Wegener's granulomatosis, cellulitis, rheumatoid arthritis, psoriatic arthritis, discoid lupus erthyematosus, systemic lupus erythematosus, scleroderma, CREST syndrome, Sjogren's syndrome, polymyositis, dermatomyositis, mixed connective tissue disease, relapsing polychondritis, vasculitis, Henoch-Schonlein syndrome, erythema nodosum, polyarteritis nodosa, temporal (giant cell) arteritis, Takayasu's arteritis, Wegener's granulomatosis, Reiter's syndrome, Behcet's syndrome, ankylosing spondylitis, cellulitis, keloids, Ehler Danlos syndrome, Marfan syndrome, pseudoxantoma elasticum, osteogenese imperfecta, chondrodysplasias, epidermolysis bullosa, Alport syndrome, and cutis laxa. TABLE 1D Gene No. cDNA Clone ID Preferred Indication Identifiers 1 H6BSF56 Cancer 2 H6EDM64 Cancer 3 H6EEC72 Cancer 4 HACAB68 Connective/Epithelial, Immune/Hematopoietic 5 HACBJ56 Cancer 6 HACBS22 Cancer 7 HADDE71 Cancer 8 HADDJ13 Connective/Epithelial 9 HADMB15 Cancer 10 HAGBQ12 Excretory, Neural/Sensory 11 HAGDW20 Neural/Sensory, Reproductive 12 HAGEG10 Cancer 13 HAGEQ79 Digestive, Neural/Sensory 14 HAGFS57 Cancer 15 HAGHN57 Cancer 16 HAHEA15 Cardiovascular 17 HAJAA47 Immune/Hematopoietic 18 HAJAY92 Cancer 19 HAJBV67 Cancer 20 HAJCH70 Cancer 21 HAOAG15 Cancer 22 HAQAI92 Digestive, Mixed Fetal, Reproductive 23 HAQCE11 Reproductive 24 HATBI94 Cancer 25 HATCB45 Cancer 26 HATCD80 Endocrine, Reproductive 27 HATCI03 Endocrine, Immune/Hematopoietic, Neural/Sensory 28 HATEH20 Cancer 29 HBAGD86 Cancer 30 HBCJL35 Cancer 31 HBDAB91 Immune/Hematopoietic 32 HBDAB91 Immune/Hematopoietic 33 HBGBC29 Cancer 34 HBGNC72 Cancer 35 HBHAA05 Neural/Sensory 36 HBHAA81 Cancer 37 HBIAA59 Cancer 38 HBIAC29 Cancer 39 HBICW51 Digestive, Immune/Hematopoietic, Neural/Sensory 40 HBJAB02 Cancer 41 HBJAC65 Cancer 42 HBJBM12 Immune/Hematopoietic 43 HBJCR46 Cancer 44 HBJDS79 Cancer 45 HBJDW56 Immune/Hematopoietic 46 HBJEL16 Cancer 47 HBJFK45 Immune/Hematopoietic 48 HBJIG20 Cancer 49 HBJKD16 Cancer 50 HBMBM96 Digestive, Immune/Hematopoietic, Neural/Sensory 51 HBMBX01 Cancer 52 HBMTM11 Cancer 53 HBMTX26 Immune/Hematopoietic 54 HBMTY48 Immune/Hematopoietic, Reproductive 55 HBMUH74 Cardiovascular, Immune/Hematopoietic, Reproductive 56 HBMWE61 Immune/Hematopoietic 57 HBNAX40 Cancer 58 HBNBJ76 Cancer 59 HBQAB79 Neural/Sensory 60 HBQAC57 Neural/Sensory 61 HBSAK32 Mixed Fetal, Musculoskeletal, Neural/Sensory 62 HBXCM66 Cardiovascular, Neural/Sensory, Reproductive 63 HBXCX15 Immune/Hematopoietic, Neural/Sensory 64 HCDCY76 Cancer 65 HCDDL48 Musculoskeletal 66 HCE1G78 Cancer 67 HCE2H52 Cancer 68 HCE3B04 Digestive, Neural/Sensory 69 HCE5F78 Immune/Hematopoietic, Neural/Sensory 70 HCEDR26 Digestive, Immune/Hematopoietic, Neural/Sensory 71 HCEEE79 Neural/Sensory 72 HCEEQ25 Mixed Fetal, Neural/Sensory 73 HCEEU18 Neural/Sensory 74 HCEFZ82 Cancer 75 HCEGX05 Cancer 76 HCFLN88 Cancer 77 HCFLT90 Cancer 78 HCHAB84 Cancer 79 HCMSX51 Cancer 80 HCNCO11 Digestive 81 HCNSD29 Cardiovascular, Digestive, Immune/Hematopoietic 82 HCQBH72 Digestive, Excretory, Immune/Hematopoietic 83 HCQCC96 Cancer 84 HCQCJ56 Cardiovascular, Digestive, Reproductive 85 HCQCM24 Cancer 86 HCRAY10 Cancer 87 HCRBF72 Cancer 88 HCRNF78 Cancer 89 HCUAF85 Immune/Hematopoietic 90 HCUCF89 Immune/Hematopoietic 91 HCUCK44 Cancer 92 HCUDD64 Cancer 93 HCWAE64 Immune/Hematopoietic 94 HCWFU39 Immune/Hematopoietic, Neural/Sensory 95 HCWUL09 Immune/Hematopoietic, Neural/Sensory 96 HDHAA42 Cancer 97 HDHEB76 Mixed Fetal, Neural/Sensory 98 HDPCW16 Cancer 99 HDPDI72 Immune/Hematopoietic 100 HDPDJ58 Cancer 101 HDPFF10 Cancer 102 HDPFU43 Cancer 103 HDPFY18 Cancer 104 HDPGE24 Cancer 105 HDPIU94 Cancer 106 HDPOC24 Cancer 107 HDPOL37 Immune/Hematopoietic, Reproductive 108 HDPOO76 Cancer 109 HDPPD93 Cancer 110 HDPPQ30 Connective/Epithelial, Immune/Hematopoietic, Musculoskeletal 111 HDPPW82 Immune/Hematopoietic 112 HDPXN20 Immune/Hematopoietic 113 HDQHM36 Immune/Hematopoietic 114 HDTAU35 Immune/Hematopoietic 115 HDTAV54 Cancer 116 HDTFX18 Immune/Hematopoietic, Reproductive 117 HDTGW48 Immune/Hematopoietic, Reproductive 118 HDTLM18 Immune/Hematopoietic 119 HE2CA60 Cancer 120 HE2CA60 Cancer 121 HE2CH58 Digestive, Mixed Fetal 122 HE2CM39 Cancer 123 HE2HC60 Cancer 124 HE2PO93 Cancer 125 HE6AU52 Mixed Fetal 126 HE6CS65 Cancer 127 HE6DO92 Immune/Hematopoietic, Mixed Fetal 128 HE6EY13 Cancer 129 HE6FU11 Mixed Fetal, Neural/Sensory, Respiratory 130 HE6FV29 Cancer 131 HE8FC45 Cancer 132 HE8FC45 Cancer 133 HE8FD92 Cancer 134 HE8FD92 Cancer 135 HE8FD92 Cancer 136 HE8FD92 Cancer 137 HE8FD92 Cancer 138 HE8SG96 Mixed Fetal, Neural/Sensory 139 HE8TY46 Cancer 140 HE9CY05 Mixed Fetal 141 HE9EA10 Cancer 142 HE9GG20 Cancer 143 HEBCI18 Cancer 144 HEBCY54 Cancer 145 HEBDF77 Neural/Sensory 146 HEBDQ91 Neural/Sensory 147 HEBFR46 Cancer 148 HEBGE07 Neural/Sensory 149 HEGAU15 Excretory, Immune/Hematopoietic, Reproductive 150 HELAT35 Cardiovascular, Mixed Fetal 151 HELBU54 Cardiovascular, Digestive 152 HELGG84 Cancer 153 HELGG84 Cancer 154 HEMEY47 Cancer 155 HEOMC46 Immune/Hematopoietic 156 HEPBA14 Reproductive 157 HEQAH80 Cancer 158 HEQBF89 Reproductive 159 HETCI16 Cancer 160 HETDW58 Cancer 161 HETEY67 Connective/Epithelial, Immune/Hematopoietic, Reproductive 162 HFCDW95 Cancer 163 HFCEI04 Neural/Sensory 164 HFCFD04 Neural/Sensory 165 HFCFE20 Cancer 166 HFEAY59 Connective/Epithelial 167 HFGAJ16 Cancer 168 HFIHZ75 Cancer 169 HFIJA29 Cancer 170 HFIJA68 Musculoskeletal 171 HFKES05 Cancer 172 HFKEU12 Excretory 173 HFPCZ55 Cancer 174 HFPDR62 Immune/Hematopoietic, Neural/Sensory 175 HFPDS07 Cancer 176 HFRAB10 Excretory, Immune/Hematopoietic, Neural/Sensory 177 HFTBM38 Cancer 178 HFTDH56 Cancer 179 HFVGK35 Cancer 180 HFVHW43 Digestive 181 HFXAV37 Immune/Hematopoietic, Neural/Sensory 182 HFXBN86 Neural/Sensory 183 HFXBT66 Neural/Sensory 184 HFXFZ46 Neural/Sensory 185 HGBER72 Cancer 186 HGBEY14 Cancer 187 HGBGN34 Cancer 188 HGBHP91 Digestive 189 HGCAC19 Cancer 190 HGCAC19 Cancer 191 HGCAC19 Cancer 192 HHEAK45 Cancer 193 HHEGS55 Immune/Hematopoietic 194 HHEOW19 Cancer 195 HHFFF87 Cancer 196 HHFFL34 Cancer 197 HHFFS40 Cancer 198 HHGCS78 Immune/Hematopoietic 199 HHGDT26 Immune/Hematopoietic, Reproductive 200 HHPFU28 Cancer 201 HHPSA85 Cancer 202 HHSBI06 Cancer 203 HHSBI65 Cancer 204 HHSDI53 Cancer 205 HHSFC09 Cancer 206 HHSGL28 Cancer 207 HILCA24 Digestive, Immune/Hematopoietic, Reproductive 208 HILCA24 Digestive, Immune/Hematopoietic, Reproductive 209 HISAT67 Cancer 210 HJBCU75 Cancer 211 HJMAA03 Cancer 212 HJMAV41 Cancer 213 HJMAY90 Cancer 214 HJPBE39 Cancer 215 HJPBK28 Cancer 216 HJPCH08 Cancer 217 HKABU43 Cancer 218 HKACI79 Cancer 219 HKAFF50 Cancer 220 HKGBF25 Cancer 221 HKIXC44 Cancer 222 HKMLK03 Digestive, Excretory, Immune/Hematopoietic 223 HKMLM95 Cancer 224 HKTAB41 Digestive, Excretory 225 HLDBG17 Cancer 226 HLDCA54 Cancer 227 HLDQU79 Cancer 228 HLDRT09 Cancer 229 HLHAP05 Immune/Hematopoietic, Neural/Sensory, Respiratory 230 HLHCS23 Respiratory 231 HLIBO72 Cancer 232 HLICE88 Cancer 233 HLICO10 Cancer 234 HLJBS28 Cancer 235 HLMBW89 Cancer 236 HLMGP50 Digestive, Immune/Hematopoietic 237 HLMJB64 Cancer 238 HLMMX62 Cancer 239 HLQAS12 Cancer 240 HLQCL64 Digestive, Immune/Hematopoietic, Reproductive 241 HLQCX36 Digestive 242 HLWAF06 Immune/Hematopoietic, Reproductive 243 HLWAU42 Cancer 244 HLWAU42 Cancer 245 HLWAV47 Cancer 246 HLWBB73 Cancer 247 HLWCN37 Cancer 248 HLWDB73 Cancer 249 HLYDF73 Immune/Hematopoietic 250 HLYEU59 Immune/Hematopoietic 251 HLYGB19 Cancer 252 HLYGE16 Cancer 253 HLYGY91 Cancer 254 HMCAZ04 Cancer 255 HMCAZ04 Cancer 256 HMCAZ04 Cancer 257 HMCAZ04 Cancer 258 HMCAZ04 Cancer 259 HMCFH60 Cancer 260 HMDAB29 Digestive, Neural/Sensory 261 HMDAD44 Connective/Epithelial, Immune/Hematopoietic, Neural/Sensory 262 HMEBB82 Cancer 263 HMEDE24 Cardiovascular 264 HMEDI90 Cardiovascular, Musculoskeletal, Neural/Sensory 265 HMELM75 Cancer 266 HMIAK10 Neural/Sensory 267 HMIBF07 Neural/Sensory 268 HMICI80 Cardiovascular, Endocrine, Neural/Sensory 269 HMICP65 Cancer 270 HMJAK70 Neural/Sensory 271 HMSBE04 Immune/Hematopoietic 272 HMSCL38 Immune/Hematopoietic, Neural/Sensory 273 HMSCR69 Cancer 274 HMSHC86 Immune/Hematopoietic 275 HMSHU20 Immune/Hematopoietic, Reproductive 276 HMSHY25 Immune/Hematopoietic 277 HMTAB77 Cancer 278 HMUAE26 Cancer 279 HMUAN45 Cancer 280 HMVBC31 Cancer 281 HMVDU15 Cancer 282 HMWBL03 Cancer 283 HMWJF53 Cancer 284 HNEAK81 Immune/Hematopoietic 285 HNECL22 Cancer 286 HNECW49 Immune/Hematopoietic 287 HNEDH88 Immune/Hematopoietic 288 HNFAC50 Cancer 289 HNFGR08 Immune/Hematopoietic 290 HNFHF34 Cancer 291 HNGAK51 Immune/Hematopoietic 292 HNGAM58 Immune/Hematopoietic 293 HNGBH53 Immune/Hematopoietic 294 HNGDQ38 Immune/Hematopoietic 295 HNGDX18 Cancer 296 HNGDY34 Immune/Hematopoietic 297 HNGEA34 Digestive, Immune/Hematopoietic 298 HNGEQ75 Immune/Hematopoietic, Neural/Sensory 299 HNGGA68 Immune/Hematopoietic, Musculoskeletal 300 HNGGP65 Immune/Hematopoietic 301 HNGHZ69 Immune/Hematopoietic 302 HNGIV64 Immune/Hematopoietic 303 HNGJB41 Immune/Hematopoietic 304 HNGKT41 Immune/Hematopoietic 305 HNGMW45 Immune/Hematopoietic 306 HNGNK44 Immune/Hematopoietic 307 HNGNO53 Immune/Hematopoietic 308 HNGPJ25 Immune/Hematopoietic, Mixed Fetal, Musculoskeletal 309 HNHEN82 Cancer 310 HNHFE71 Immune/Hematopoietic 311 HNHGK22 Immune/Hematopoietic 312 HNHHB10 Immune/Hematopoietic, Reproductive 313 HNHKS19 Immune/Hematopoietic, Reproductive 314 HNTBT17 Cancer 315 HNTMH79 Cancer 316 HOABP31 Cancer 317 HOABP31 Cancer 318 HOACG07 Cancer 319 HODAG07 Reproductive 320 HODBB70 Reproductive 321 HODBV05 Cancer 322 HODCZ32 Reproductive 323 HOEBK60 Cancer 324 HOFAA78 Cancer 325 HOFNB74 Reproductive 326 HOFNU55 Reproductive 327 HOGBF01 Reproductive 328 HORBS82 Cancer 329 HORBV76 Immune/Hematopoietic, Reproductive 330 HOSDO75 Cancer 331 HOSEC25 Musculoskeletal 332 HOSEI81 Digestive, Musculoskeletal 333 HOSEJ94 Cancer 334 HOUCA21 Connective/Epithelial, Immune/Hematopoietic, Musculoskeletal 335 HOUDE92 Cancer 336 HOUDR07 Cancer 337 HOUED72 Connective/Epithelial 338 HOUFS04 Cancer 339 HOUHI25 Cancer 340 HOVBD85 Musculoskeletal, Reproductive 341 HPCAB41 Immune/Hematopoietic, Reproductive 342 HPCAL26 Cancer 343 HPEAD23 Cancer 344 HPFBA54 Reproductive 345 HPFCI36 Cancer 346 HPFDI37 Cancer 347 HPIAA80 Cancer 348 HPJBJ51 Immune/Hematopoietic, Reproductive 349 HPJBJ51 Immune/Hematopoietic, Reproductive 350 HPJBU43 Reproductive 351 HPJCW58 Reproductive 352 HPMBX22 Cancer 353 HPMCJ84 Reproductive 354 HPMCV30 Cancer 355 HPMFH77 Cancer 356 HPQAX38 Cardiovascular 357 HPQAX38 Cardiovascular 358 HPQCB83 Cancer 359 HPQCC53 Cancer 360 HPRBH85 Cancer 361 HPRCA64 Cancer 362 HPRCD35 Cancer 363 HPTRM02 Cancer 364 HPWBA29 Reproductive 365 HPWDK06 Cancer 366 HRAAD30 Cancer 367 HRADA42 Cancer 368 HRADF49 Cancer 369 HRADN25 Cancer 370 HRADT25 Digestive, Excretory 371 HRDAI17 Cancer 372 HRDDQ39 Cancer 373 HRDER22 Cancer 374 HRDEX93 Cancer 375 HRDFK37 Cancer 376 HRGBD54 Cancer 377 HROEA08 Cancer 378 HSAVA08 Immune/Hematopoietic 379 HSAVW42 Cancer 380 HSAWN53 Immune/Hematopoietic 381 HSAWZ40 Immune/Hematopoietic 382 HSAYC41 Excretory, Immune/Hematopoietic, Reproductive 383 HSDZM54 Neural/Sensory 384 HSHBF76 Cancer 385 HSIFG47 Digestive 386 HSJBY32 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 387 HSKDR27 Cancer 388 HSLHG78 Cancer 389 HSLHX15 Musculoskeletal 390 HSNAP85 Cancer 391 HSNAZ09 Cancer 392 HSNBM34 Cancer 393 HSOAH16 Digestive 394 HSQBF66 Cancer 395 HSQDO85 Cancer 396 HSQES57 Cancer 397 HSRBE06 Cancer 398 HSSDI26 Musculoskeletal 399 HSSEA64 Cancer 400 HSSEF77 Cancer 401 HSSFE38 Cancer 402 HSSGJ58 Musculoskeletal 403 HSWBE76 Cancer 404 HSXCP38 Cardiovascular, Neural/Sensory 405 HSYBI06 Cancer 406 HT1SC27 Digestive, Immune/Hematopoietic, Reproductive 407 HT3BF49 Immune/Hematopoietic 408 HT4FV41 Cancer 409 HT5FX79 Cancer 410 HT5GR59 Cancer 411 HTAEI78 Immune/Hematopoietic 412 HTDAA78 Immune/Hematopoietic 413 HTEAG62 Digestive, Immune/Hematopoietic, Reproductive 414 HTECB02 Cancer 415 HTECC15 Cancer 416 HTEDF18 Reproductive 417 HTEDJ28 Cancer 418 HTEDS12 Cardiovascular, Immune/Hematopoietic, Reproductive 419 HTEED26 Reproductive 420 HTEED26 Reproductive 421 HTEEF26 Cancer 422 HTEEF26 Cancer 423 HTEEW69 Reproductive 424 HTEGS07 Reproductive 425 HTEGS11 Cancer 426 HTEHA56 Cancer 427 HTEHU59 Cancer 428 HTEJD29 Reproductive 429 HTEKM46 Cancer 430 HTEMQ17 Cancer 431 HTENR63 Cancer 432 HTGGM44 Cancer 433 HTHBZ06 Cancer 434 HTLAP64 Cancer 435 HTLBT80 Cancer 436 HTLDA84 Reproductive 437 HTLDN29 Cancer 438 HTLDU78 Reproductive 439 HTLEC82 Cancer 440 HTLEM16 Cancer 441 HTLEV48 Reproductive 442 HTLFA13 Musculoskeletal, Reproductive 443 HTLFI73 Digestive, Immune/Hematopoietic, Reproductive 444 HTLGI89 Cancer 445 HTLIF11 Cancer 446 HTLIF12 Excretory, Reproductive 447 HTLIF12 Excretory, Reproductive 448 HTLIF12 Excretory, Reproductive 449 HTLIF12 Excretory, Reproductive 450 HTLIF12 Excretory, Reproductive 451 HTLIF12 Excretory, Reproductive 452 HTNAM63 Endocrine 453 HTNBK13 Cancer 454 HTOAI50 Digestive, Immune/Hematopoietic 455 HTOAM11 Immune/Hematopoietic, Neural/Sensory 456 HTODH57 Immune/Hematopoietic 457 HTODH83 Immune/Hematopoietic 458 HTOEV16 Cancer 459 HTOGR38 Immune/Hematopoietic 460 HTOHO21 Immune/Hematopoietic 461 HTOHQ05 Immune/Hematopoietic 462 HTOJL95 Cancer 463 HTOJL95 Cancer 464 HTPDU17 Cancer 465 HTSFJ32 Immune/Hematopoietic 466 HTTCB60 Cancer 467 HTTEE41 Cancer 468 HTTEZ02 Cancer 469 HTWEH94 Immune/Hematopoietic 470 HTXBD09 Cancer 471 HTXDB22 Cancer 472 HTXDC38 Cancer 473 HTXDC77 Cancer 474 HTXDD61 Cancer 475 HTXDG92 Cancer 476 HTXET11 Digestive, Immune/Hematopoietic 477 HTXFA72 Immune/Hematopoietic 478 HTXJY08 Cancer 479 HTXKF95 Cancer 480 HTXMZ07 Cancer 481 HUFCL31 Digestive, Immune/Hematopoietic 482 HUKBT67 Cancer 483 HUKDF20 Cardiovascular, Neural/Sensory, Reproductive 484 HUKDY82 Cancer 485 HUSCJ14 Cancer 486 HUSGL67 Cancer 487 HUSGU40 Cancer 488 HUSIR18 Cancer 489 HUVDJ48 Digestive, Reproductive 490 HWAAI12 Cancer 491 HWBBQ70 Immune/Hematopoietic, Neural/Sensory 492 HWBCN36 Immune/Hematopoietic 493 HWBDJ08 Cancer 494 HWBFX16 Immune/Hematopoietic 495 HWDAC26 Connective/Epithelial, Immune/Hematopoietic, Neural/Sensory 496 HWDAG96 Cancer 497 HWDAJ01 Connective/Epithelial 498 HWHPB78 Cancer 499 HYABC84 Cancer 500 HYABC84 Cancer 501 H2CBD20 Digestive 502 H2CBH91 Cancer 503 H2LBA54 Cancer 504 H2LBB09 Cancer 505 H2LBB09 Cancer 506 H2MAC63 Digestive, Reproductive 507 H2MBA76 Cancer 508 H2MBF60 Cancer 509 H6BSM88 Cancer 510 H6EEA48 Cancer 511 H6EEN71 Cancer 512 H6EEO05 Cancer 513 H6EEU40 Cancer 514 H7TDB54 Cancer 515 H7TMB95 Cancer 516 HAAAT06 Cancer 517 HACAD42 Connective/Epithelial, Mixed Fetal, Neural/Sensory 518 HACBJ11 Cancer 519 HACBS86 Cancer 520 HACBT91 Cancer 521 HACBZ73 Cancer 522 HACCK29 Connective/Epithelial 523 HADAB60 Cancer 524 HADAM31 Connective/Epithelial 525 HADCL19 Connective/Epithelial 526 HADCZ65 Connective/Epithelial, Immune/Hematopoietic 527 HADDC04 Connective/Epithelial, Reproductive 528 HADDP23 Cancer 529 HADDP51 Cancer 530 HADDR24 Cancer 531 HADET62 Connective/Epithelial 532 HADEY08 Cancer 533 HADEY22 Connective/Epithelial 534 HADEY22 Connective/Epithelial 535 HADFB84 Cancer 536 HADFD01 Cancer 537 HADFD10 Cancer 538 HADFK11 Connective/Epithelial 539 HADFT44 Connective/Epithelial, Mixed Fetal, Neural/Sensory 540 HADFW20 Connective/Epithelial 541 HADFX10 Connective/Epithelial, Neural/Sensory 542 HADFY80 Connective/Epithelial, Digestive 543 HADGD93 Cardiovascular, Connective/Epithelial 544 HADMA77 Cancer 545 HADXA10 Cancer 546 HADXA10 Cancer 547 HAFBB15 Cancer 548 HAFBL14 Cancer 549 HAGAB62 Cancer 550 HAGAB83 Neural/Sensory 551 HAGAE84 Neural/Sensory 552 HAGAF75 Digestive, Neural/Sensory 553 HAGAK40 Cancer 554 HAGAU43 Neural/Sensory 555 HAGAZ36 Neural/Sensory 556 HAGBC57 Cancer 557 HAGBL31 Neural/Sensory 558 HAGBO09 Mixed Fetal, Neural/Sensory 559 HAGBO12 Neural/Sensory 560 HAGBO51 Neural/Sensory 561 HAGBS89 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 562 HAGBV06 Cancer 563 HAGBV25 Cancer 564 HAGBV29 Immune/Hematopoietic, Neural/Sensory 565 HAGCC87 Digestive, Immune/Hematopoietic, Neural/Sensory 566 HAGCH67 Neural/Sensory 567 HAGCI69 Neural/Sensory, Reproductive 568 HAGCT33 Immune/Hematopoietic, Mixed Fetal, Neural/Sensory 569 HAGCZ70 Neural/Sensory 570 HAGDC73 Cancer 571 HAGDG84 Immune/Hematopoietic, Neural/Sensory 572 HAGDH85 Neural/Sensory 573 HAGDI69 Neural/Sensory 574 HAGDJ53 Immune/Hematopoietic, Neural/Sensory 575 HAGDJ56 Cardiovascular, Endocrine, Neural/Sensory 576 HAGDL51 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 577 HAGDO70 Cancer 578 HAGDT30 Cancer 579 HAGDW68 Endocrine, Neural/Sensory 580 HAGDX84 Cancer 581 HAGEK37 Cancer 582 HAGEK86 Cancer 583 HAGEP30 Neural/Sensory 584 HAGEQ58 Neural/Sensory 585 HAGEQ67 Cancer 586 HAGEU26 Neural/Sensory 587 HAGEW83 Neural/Sensory 588 HAGEX49 Cancer 589 HAGEX49 Cancer 590 HAGFD75 Cancer 591 HAGFF43 Cancer 592 HAGFJ67 Digestive, Immune/Hematopoietic, Neural/Sensory 593 HAGFM58 Cardiovascular, Neural/Sensory 594 HAGFT48 Cancer 595 HAGFU31 Neural/Sensory 596 HAGFW13 Neural/Sensory 597 HAGHE85 Cardiovascular, Neural/Sensory 598 HAGHR18 Neural/Sensory 599 HAGIB90 Cancer 600 HAHEM51 Cardiovascular 601 HAHSA76 Cardiovascular 602 HAHSD51 Cancer 603 HAIBR76 Cancer 604 HAIBT20 Cancer 605 HAIBV91 Cancer 606 HAICE62 Cancer 607 HAICL90 Digestive, Immune/Hematopoietic, Reproductive 608 HAICV44 Cancer 609 HAIDP45 Cancer 610 HAJAB88 Cancer 611 HAJAZ56 Cancer 612 HAMFC67 Cancer 613 HAMFQ38 Cancer 614 HANGG01 Cancer 615 HANGB24 Cancer 616 HANKC93 Musculoskeletal 617 HAPAD35 Cancer 618 HAPBR13 Cancer 619 HAPBU09 Cancer 620 HAPBU86 Cancer 621 HAPBU86 Cancer 622 HAPNJ33 Cancer 623 HAPNL62 Cancer 624 HAPNO50 Cancer 625 HAPNY10 Cancer 626 HAPPW83 Cancer 627 HAPQJ73 Cancer 628 HAPQK26 Reproductive 629 HAPQU71 Cancer 630 HAPQU71 Cancer 631 HAPQW18 Cancer 632 HAPQX44 Cancer 633 HAPRK55 Cancer 634 HAPSH37 Cancer 635 HAQBG57 Cancer 636 HAQBY85 Cancer 637 HAQBZ15 Cancer 638 HAQCE18 Immune/Hematopoietic, Reproductive 639 HAQCF94 Cancer 640 HARAE26 Neural/Sensory 641 HARAT69 Cancer 642 HARAZ81 Cancer 643 HASAU26 Cancer 644 HASAX57 Cancer 645 HASAY07 Cancer 646 HATAE01 Cancer 647 HATAG52 Endocrine, Neural/Sensory, Reproductive 648 HATAL05 Cancer 649 HATBA90 Endocrine 650 HATBM71 Endocrine 651 HATCF80 Cancer 652 HATCI67 Cancer 653 HATCJ27 Cancer 654 HATCS79 Endocrine, Immune/Hematopoietic 655 HATCX03 Cancer 656 HATDE03 Cancer 657 HATDF41 Cancer 658 HATDH23 Cancer 659 HATDH55 Cancer 660 HATDO84 Endocrine 661 HATDU01 Cancer 662 HATDW05 Endocrine 663 HATEF13 Digestive, Endocrine 664 HATEF64 Cancer 665 HATEH40 Cancer 666 HATEI22 Cancer 667 HAUCC84 Cancer 668 HAWAS41 Connective/Epithelial, Excretory, Immune/Hematopoietic 669 HAWBA65 Cancer 670 HBAGH64 Cancer 671 HBAGV01 Connective/Epithelial, Excretory 672 HBAMC50 Excretory 673 HBAMC57 Excretory 674 HBBBA42 Cancer 675 HBBBB08 Neural/Sensory 676 HBBBE83 Cancer 677 HBBMA11 Neural/Sensory 678 HBCAK10 Digestive, Immune/Hematopoietic, Reproductive 679 HBCAK80 Cancer 680 HBCAQ48 Cancer 681 HBCAY17 Cancer 682 HBCGE46 Musculoskeletal 683 HBGBA14 Cancer 684 HBGBE75 Cancer 685 HBGBP22 Cancer 686 HBGFQ34 Reproductive 687 HBGML95 Reproductive 688 HBGMT60 Cancer 689 HBHAA53 Neural/Sensory 690 HBIAU43 Cancer 691 HBIAW58 Neural/Sensory 692 HBIBB20 Cancer 693 HBIBF26 Cancer 694 HBIBM33 Neural/Sensory 695 HBIBN67 Cancer 696 HBIBQ69 Immune/Hematopoietic, Neural/Sensory 697 HBIBR38 Neural/Sensory 698 HBIBR61 Cancer 699 HBIBS33 Neural/Sensory 700 HBIBT13 Digestive, Immune/Hematopoietic, Neural/Sensory 701 HBIBZ20 Neural/Sensory 702 HBICB80 Cancer 703 HBJAC40 Cancer 704 HBJAV56 Immune/Hematopoietic, Musculoskeletal 705 HBJAY14 Immune/Hematopoietic 706 HBJBQ69 Immune/Hematopoietic 707 HBJBR40 Immune/Hematopoietic 708 HBJCH46 Immune/Hematopoietic, Musculoskeletal 709 HBJCR17 Cancer 710 HBJCS26 Cancer 711 HBJCW24 Cancer 712 HBJDC57 Immune/Hematopoietic, Reproductive 713 HBJDR18 Immune/Hematopoietic 714 HBJDR83 Immune/Hematopoietic 715 HBJEE51 Immune/Hematopoietic 716 HBJEL21 Cancer 717 HBJFH84 Cancer 718 HBJFJ14 Cancer 719 HBJFJ26 Cancer 720 HBJFJ83 Immune/Hematopoietic, Mixed Fetal 721 HBJFJ83 Immune/Hematopoietic, Mixed Fetal 722 HBJFP47 Immune/Hematopoietic, Reproductive 723 HBJFR77 Cancer 724 HBJFU30 Cancer 725 HBJFX41 Immune/Hematopoietic 726 HBJHO83 Immune/Hematopoietic, Reproductive 727 HBJHS92 Immune/Hematopoietic, Neural/Sensory 728 HBJHT01 Immune/Hematopoietic, Reproductive 729 HBJHT01 Immune/Hematopoietic, Reproductive 730 HBJHW06 Immune/Hematopoietic, Reproductive 731 HBJIR14 Cancer 732 HBJJA26 Immune/Hematopoietic 733 HBJJX02 Immune/Hematopoietic 734 HBJLH78 Immune/Hematopoietic 735 HBJND04 Cancer 736 HBJND57 Immune/Hematopoietic 737 HBKDF66 Cancer 738 HBKEA94 Cancer 739 HBKEE60 Digestive 740 HBKEI41 Endocrine, Mixed Fetal, Reproductive 741 HBMBD51 Digestive, Immune/Hematopoietic 742 HBMBD73 Cancer 743 HBMBE33 Immune/Hematopoietic 744 HBMBM17 Immune/Hematopoietic, Reproductive 745 HBMCL59 Immune/Hematopoietic 746 HBMCM96 Immune/Hematopoietic, Neural/Sensory 747 HBMCQ74 Cancer 748 HBMCQ74 Cancer 749 HBMCT40 Cancer 750 HBMDM08 Immune/Hematopoietic 751 HBMSN62 Cancer 752 HBMSO30 Immune/Hematopoietic 753 HBMTM50 Cancer 754 HBMUD59 Cancer 755 HBMUI10 Cancer 756 HBMUJ48 Cancer 757 HBMUR39 Immune/Hematopoietic 758 HBMVF65 Endocrine, Immune/Hematopoietic, Neural/Sensory 759 HBMVF65 Endocrine, Immune/Hematopoietic, Neural/Sensory 760 HBMWC39 Cancer 761 HBMWJ92 Cancer 762 HBMWS52 Immune/Hematopoietic 763 HBMXE34 Cancer 764 HBMXG01 Immune/Hematopoietic 765 HBMXG76 Immune/Hematopoietic 766 HBMXM05 Excretory, Immune/Hematopoietic, Neural/Sensory 767 HBMXW83 Cancer 768 HBNAE74 Excretory, Musculoskeletal, Reproductive 769 HBNAX16 Cancer 770 HBNAZ35 Endocrine, Reproductive 771 HBODK40 Cancer 772 HBODV76 Cancer 773 HBPAD89 Cancer 774 HBPAF39 Immune/Hematopoietic, Neural/Sensory 775 HBQAC45 Neural/Sensory 776 HBQAC72 Neural/Sensory 777 HBQAE37 Neural/Sensory 778 HBSAJ63 Cancer 779 HBSAJ63 Cancer 780 HBSDD24 Cancer 781 HBWBD25 Immune/Hematopoietic, Neural/Sensory 782 HBXAS93 Neural/Sensory 783 HBXAT27 Cancer 784 HBXAW57 Neural/Sensory 785 HBXBI29 Neural/Sensory 786 HBXBM24 Neural/Sensory 787 HBXBM78 Cancer 788 HBXCD59 Immune/Hematopoietic, Neural/Sensory 789 HBXCE43 Neural/Sensory 790 HBXCG08 Cancer 791 HBXCM52 Cancer 792 HBXCQ03 Cancer 793 HBXCR15 Cancer 794 HBXDL52 Cancer 795 HBXDL52 Cancer 796 HBXDN08 Cancer 797 HBXDN65 Neural/Sensory 798 HBXFA04 Neural/Sensory 799 HBXFE64 Neural/Sensory 800 HBXFI33 Immune/Hematopoietic, Neural/Sensory 801 HBXFP72 Cancer 802 HBXFS31 Neural/Sensory 803 HBXFW01 Neural/Sensory 804 HBXGE12 Cancer 805 HBXGL91 Neural/Sensory, Reproductive 806 HBXGM24 Cancer 807 HBZAI75 Digestive, Reproductive 808 HCABP33 Cancer 809 HCABW10 Cancer 810 HCACZ65 Cancer 811 HCBAB34 Cancer 812 HCDAA24 Cancer 813 HCDAA24 Cancer 814 HCDAF17 Cancer 815 HCDAH02 Immune/Hematopoietic, Musculoskeletal 816 HCDAP33 Cancer 817 HCDAR40 Cardiovascular, Immune/Hematopoietic, Musculoskeletal 818 HCDAS02 Cancer 819 HCDBE76 Cancer 820 HCDBO32 Cancer 821 HCDBW67 Cancer 822 HCDBZ31 Musculoskeletal 823 HCDCB03 Cancer 824 HCDCE51 Cancer 825 HCDCI42 Immune/Hematopoietic, Musculoskeletal 826 HCDDB15 Cancer 827 HCDDX81 Musculoskeletal 828 HCDDY28 Cardiovascular, Musculoskeletal 829 HCDEB19 Cancer 830 HCDEN46 Cancer 831 HCDES69 Immune/Hematopoietic, Musculoskeletal, Reproductive 832 HCE1D45 Cancer 833 HCE1N56 Cancer 834 HCE1T53 Neural/Sensory 835 HCE1Y27 Digestive, Neural/Sensory, Reproductive 836 HCE1Y34 Immune/Hematopoietic, Neural/Sensory 837 HCE2B57 Musculoskeletal, Neural/Sensory 838 HCE2E47 Immune/Hematopoietic, Mixed Fetal, Neural/Sensory 839 HCE2I23 Neural/Sensory 840 HCE2P90 Neural/Sensory 841 HCE3A54 Neural/Sensory 842 HCE3C46 Immune/Hematopoietic, Neural/Sensory 843 HCE3D58 Cancer 844 HCE3D89 Endocrine, Neural/Sensory 845 HCE3J43 Cancer 846 HCE3L04 Neural/Sensory 847 HCE3N23 Cancer 848 HCE3R01 Cancer 849 HCE3R01 Cancer 850 HCE3R01 Cancer 851 HCE3R46 Cancer 852 HCE4H32 Cancer 853 HCE4H32 Cancer 854 HCE4T64 Cancer 855 HCE4W88 Cancer 856 HCE5B62 Neural/Sensory 857 HCE5H86 Cancer 858 HCE5J64 Digestive, Neural/Sensory 859 HCEBF54 Cancer 860 HCECO77 Cancer 861 HCEDH42 Neural/Sensory 862 HCEDJ05 Neural/Sensory 863 HCEDJ26 Cancer 864 HCEDN07 Digestive, Mixed Fetal, Neural/Sensory 865 HCEDO17 Cancer 866 HCEEG48 Neural/Sensory 867 HCEEM33 Cancer 868 HCEEP16 Immune/Hematopoietic, Neural/Sensory, Reproductive 869 HCEER60 Cardiovascular, Digestive, Neural/Sensory 870 HCEFA10 Immune/Hematopoietic, Neural/Sensory, Reproductive 871 HCEFA50 Neural/Sensory 872 HCEFA94 Neural/Sensory 873 HCEFC27 Cancer 874 HCEFG93 Neural/Sensory 875 HCEFH31 Cancer 876 HCEFK56 Cancer 877 HCEFN51 Cancer 878 HCEGG08 Cancer 879 HCEGH74 Cancer 880 HCEGK81 Cancer 881 HCEGS49 Connective/Epithelial, Neural/Sensory, Reproductive 882 HCEGU75 Cancer 883 HCEGY33 Cancer 884 HCEHW24 Neural/Sensory 885 HCEJL08 Cancer 886 HCEJP93 Cancer 887 HCELB04 Cancer 888 HCEMA08 Cancer 889 HCENN67 Digestive, Endocrine, Neural/Sensory 890 HCENQ22 Digestive, Immune/Hematopoietic, Neural/Sensory 891 HCEOF01 Neural/Sensory, Reproductive 892 HCEOF01 Neural/Sensory, Reproductive 893 HCEON94 Immune/Hematopoietic, Neural/Sensory, Reproductive 894 HCEOQ67 Cancer 895 HCEOV48 Cancer 896 HCEPC90 Neural/Sensory 897 HCEPO08 Cancer 898 HCESB03 Immune/Hematopoietic, Neural/Sensory 899 HCESK44 Cancer 900 HCETE08 Mixed Fetal, Neural/Sensory, Reproductive 901 HCETL19 Immune/Hematopoietic, Neural/Sensory, Reproductive 902 HCEWD90 Cancer 903 HCEWE62 Neural/Sensory 904 HCEZW14 Cancer 905 HCFAT42 Immune/Hematopoietic 906 HCFAT66 Immune/Hematopoietic 907 HCFBA30 Cardiovascular, Immune/Hematopoietic, Reproductive 908 HCFBM77 Immune/Hematopoietic 909 HCFBV39 Cancer 910 HCFCB72 Immune/Hematopoietic 911 HCFCG91 Cancer 912 HCFCM81 Digestive, Immune/Hematopoietic, Reproductive 913 HCFCW39 Cancer 914 HCFCY49 Cancer 915 HCFDD18 Digestive, Immune/Hematopoietic, Mixed Fetal 916 HCFLB10 Cardiovascular, Immune/Hematopoietic 917 HCFLC03 Cancer 918 HCFLJ52 Cancer 919 HCFLL33 Immune/Hematopoietic, Reproductive 920 HCFLP48 Immune/Hematopoietic 921 HCFLQ12 Cancer 922 HCFLY20 Cancer 923 HCFLY20 Cancer 924 HCFMA39 Immune/Hematopoietic 925 HCFMJ40 Immune/Hematopoietic 926 HCFML07 Cancer 927 HCFMR75 Digestive, Immune/Hematopoietic 928 HCFMX16 Immune/Hematopoietic 929 HCFMX88 Immune/Hematopoietic, Neural/Sensory 930 HCFNM40 Digestive, Immune/Hematopoietic, Reproductive 931 HCFNM50 Immune/Hematopoietic 932 HCFNN16 Cancer 933 HCFNN75 Cancer 934 HCFOG17 Immune/Hematopoietic 935 HCFOH93 Immune/Hematopoietic, Reproductive 936 HCGBA15 Cancer 937 HCHAC68 Cancer 938 HCHBP49 Cancer 939 HCHCA79 Digestive, Neural/Sensory, Reproductive 940 HCHCG33 Cancer 941 HCHMY57 Cancer 942 HCHOC06 Reproductive 943 HCHOY52 Cancer 944 HCHQB93 Cancer 945 HCHQB93 Cancer 946 HCLBK61 Cancer 947 HCLCU75 Respiratory 948 HCMSA37 Cardiovascular 949 HCMSR07 Cardiovascular 950 HCNAI74 Digestive 951 HCNCT01 Digestive 952 HCNDR39 Cancer 953 HCNSD91 Cancer 954 HCNSF01 Cancer 955 HCNSG06 Digestive, Reproductive 956 HCNSG32 Digestive, Reproductive 957 HCPAE41 Cancer 958 HCQAK36 Cancer 959 HCQAQ47 Cancer 960 HCQAS72 Cancer 961 HCQBM95 Digestive, Immune/Hematopoietic 962 HCQCM95 Cancer 963 HCQCM95 Cancer 964 HCQCV23 Cancer 965 HCQCV23 Cancer 966 HCQDD32 Digestive, Immune/Hematopoietic, Reproductive 967 HCQDD61 Digestive, Immune/Hematopoietic 968 HCQDT67 Digestive, Immune/Hematopoietic 969 HCRAI29 Neural/Sensory 970 HCRBI79 Cancer 971 HCRBL20 Cancer 972 HCRBX84 Cancer 973 HCRMA24 Digestive, Musculoskeletal 974 HCRMR35 Cancer 975 HCRMR35 Cancer 976 HCRMR35 Cancer 977 HCROC18 Cancer 978 HCUAE53 Immune/Hematopoietic 979 HCUAO46 Immune/Hematopoietic 980 HCUAT74 Cancer 981 HCUBA28 Cancer 982 HCUBC45 Cancer 983 HCUBM41 Immune/Hematopoietic 984 HCUBN69 Immune/Hematopoietic 985 HCUBY47 Digestive, Immune/Hematopoietic 986 HCUCV66 Cancer 987 HCUDJ41 Immune/Hematopoietic 988 HCUEC55 Immune/Hematopoietic 989 HCUEG85 Immune/Hematopoietic 990 HCUES35 Immune/Hematopoietic, Neural/Sensory 991 HCUFC77 Cancer 992 HCUFD17 Cancer 993 HCUFD46 Immune/Hematopoietic 994 HCUFE33 Immune/Hematopoietic 995 HCUFJ09 Cancer 996 HCUFQ58 Cardiovascular, Immune/Hematopoietic 997 HCUFQ58 Cardiovascular, Immune/Hematopoietic 998 HCUFX08 Immune/Hematopoietic 999 HCUGB76 Immune/Hematopoietic, Reproductive 1000 HCUGK79 Immune/Hematopoietic 1001 HCUGQ19 Immune/Hematopoietic 1002 HCUGR26 Immune/Hematopoietic 1003 HCUGR86 Immune/Hematopoietic 1004 HCUHE27 Immune/Hematopoietic 1005 HCUHL82 Cardiovascular, Immune/Hematopoietic, Reproductive 1006 HCUHM71 Immune/Hematopoietic, Musculoskeletal 1007 HCWAK88 Immune/Hematopoietic 1008 HCWAL10 Cardiovascular, Immune/Hematopoietic 1009 HCWAT71 Immune/Hematopoietic 1010 HCWBQ52 Immune/Hematopoietic 1011 HCWCH16 Immune/Hematopoietic 1012 HCWDM69 Digestive, Immune/Hematopoietic 1013 HCWEB38 Immune/Hematopoietic 1014 HCWEB72 Immune/Hematopoietic 1015 HCWEF04 Cancer 1016 HCWEI82 Immune/Hematopoietic 1017 HCWEM96 Cancer 1018 HCWFJ16 Immune/Hematopoietic 1019 HCWFJ16 Immune/Hematopoietic 1020 HCWFK03 Cancer 1021 HCWHD30 Immune/Hematopoietic 1022 HCWHT34 Immune/Hematopoietic, Mixed Fetal 1023 HCWHT52 Immune/Hematopoietic 1024 HCWKO32 Immune/Hematopoietic 1025 HCWLE50 Immune/Hematopoietic 1026 HCWUF93 Cancer 1027 HCWUW24 Immune/Hematopoietic 1028 HCYBA32 Cancer 1029 HDAAV67 Musculoskeletal, Neural/Sensory 1030 HDABR74 Cancer 1031 HDABW45 Immune/Hematopoietic, Musculoskeletal 1032 HDACJ52 Cancer 1033 HDCBM09 Immune/Hematopoietic 1034 HDFAB86 Mixed Fetal, Neural/Sensory 1035 HDFIB37 Connective/Epithelial, Neural/Sensory 1036 HDFMB91 Neural/Sensory 1037 HDHAA55 Immune/Hematopoietic, Neural/Sensory 1038 HDHEA33 Cancer 1039 HDHEB12 Immune/Hematopoietic, Neural/Sensory 1040 HDHEB80 Neural/Sensory 1041 HDHIA16 Cancer 1042 HDHIA26 Neural/Sensory 1043 HDHMA71 Cancer 1044 HDLAL94 Cancer 1045 HDPAB86 Cancer 1046 HDPAE80 Cancer 1047 HDPAQ86 Digestive, Immune/Hematopoietic, Neural/Sensory 1048 HDPBD56 Cancer 1049 HDPBN48 Digestive, Immune/Hematopoietic 1050 HDPCG79 Digestive, Immune/Hematopoietic, Reproductive 1051 HDPCV29 Immune/Hematopoietic 1052 HDPDA36 Immune/Hematopoietic 1053 HDPDC59 Immune/Hematopoietic, Musculoskeletal 1054 HDPFG13 Cancer 1055 HDPFK27 Immune/Hematopoietic, Neural/Sensory 1056 HDPFZ05 Immune/Hematopoietic, Neural/Sensory 1057 HDPGA84 Cancer 1058 HDPGR80 Cancer 1059 HDPGU14 Cancer 1060 HDPGX09 Cancer 1061 HDPIE44 Cancer 1062 HDPIE73 Immune/Hematopoietic 1063 HDPIF35 Immune/Hematopoietic 1064 HDPIF65 Immune/Hematopoietic 1065 HDPIH25 Cancer 1066 HDPIY31 Cancer 1067 HDPJH72 Cancer 1068 HDPJV53 Immune/Hematopoietic 1069 HDPJV75 Cancer 1070 HDPKC55 Cardiovascular, Immune/Hematopoietic, Reproductive 1071 HDPKD16 Cancer 1072 HDPMC52 Digestive, Immune/Hematopoietic, Musculoskeletal 1073 HDPML04 Connective/Epithelial, Immune/Hematopoietic 1074 HDPMM22 Immune/Hematopoietic 1075 HDPNC21 Cancer 1076 HDPNJ26 Cancer 1077 HDPOD73 Immune/Hematopoietic 1078 HDPOT33 Cancer 1079 HDPPB70 Cancer 1080 HDPPC19 Immune/Hematopoietic 1081 HDPPE05 Cancer 1082 HDPSA70 Cancer 1083 HDPSS56 Cancer 1084 HDPSZ07 Immune/Hematopoietic 1085 HDPSZ07 Immune/Hematopoietic 1086 HDPSZ07 Immune/Hematopoietic 1087 HDPTI49 Immune/Hematopoietic, Neural/Sensory 1088 HDPTP22 Immune/Hematopoietic, Neural/Sensory, Reproductive 1089 HDPYE25 Immune/Hematopoietic, Neural/Sensory 1090 HDQGD06 Cancer 1091 HDQGD06 Cancer 1092 HDQGD06 Cancer 1093 HDQGN08 Immune/Hematopoietic 1094 HDQGO62 Cancer 1095 HDQPM16 Cancer 1096 HDRAA17 Cancer 1097 HDRAC68 Cancer 1098 HDSAC78 Cancer 1099 HDSAH37 Connective/Epithelial 1100 HDSAM57 Immune/Hematopoietic 1101 HDSAO14 Cancer 1102 HDSAO64 Cancer 1103 HDSAP15 Cancer 1104 HDTAR39 Cancer 1105 HDTAS57 Cancer 1106 HDTBP62 Cancer 1107 HDTBQ77 Cancer 1108 HDTDA48 Immune/Hematopoietic, Neural/Sensory 1109 HDTDE66 Cancer 1110 HDTDG75 Immune/Hematopoietic 1111 HDTDS09 Cancer 1112 HDTFF53 Cancer 1113 HDTGW76 Cancer 1114 HDTGZ56 Immune/Hematopoietic 1115 HDTHZ85 Cancer 1116 HDTIM39 Cancer 1117 HDTKJ29 Cancer 1118 HDUAB12 Cancer 1119 HDUAD68 Cancer 1120 HE2AC74 Cancer 1121 HE2AC74 Cancer 1122 HE2AC75 Mixed Fetal 1123 HE2AI94 Cancer 1124 HE2AT61 Cancer 1125 HE2AX36 Cancer 1126 HE2AY96 Cancer 1127 HE2BD72 Cancer 1128 HE2BH50 Cancer 1129 HE2CB53 Cancer 1130 HE2CC17 Excretory, Mixed Fetal 1131 HE2CJ53 Cancer 1132 HE2CK47 Cancer 1133 HE2CM34 Cancer 1134 HE2DG46 Mixed Fetal 1135 HE2DI16 Mixed Fetal 1136 HE2DJ84 Cancer 1137 HE2DY23 Cancer 1138 HE2DY25 Cancer 1139 HE2EE80 Cancer 1140 HE2EH45 Mixed Fetal 1141 HE2FE89 Cardiovascular, Digestive, Mixed Fetal 1142 HE2FR49 Cancer 1143 HE2GB19 Cancer 1144 HE2GO81 Cancer 1145 HE2HB61 Mixed Fetal 1146 HE2HB64 Cancer 1147 HE2HF76 Cancer 1148 HE2ID09 Mixed Fetal 1149 HE2IE66 Cancer 1150 HE2NW57 Mixed Fetal 1151 HE2OA95 Cancer 1152 HE2OC39 Mixed Fetal, Musculoskeletal 1153 HE2PB61 Cancer 1154 HE2PI43 Cancer 1155 HE2PJ56 Cancer 1156 HE6CJ41 Immune/Hematopoietic, Mixed Fetal 1157 HE6DC37 Digestive, Mixed Fetal, Reproductive 1158 HE6DN83 Cancer 1159 HE6EI30 Immune/Hematopoietic, Mixed Fetal 1160 HE6ET70 Mixed Fetal, Reproductive 1161 HE6GO65 Mixed Fetal, Neural/Sensory 1162 HE8AN83 Mixed Fetal, Musculoskeletal 1163 HE8AU68 Cancer 1164 HE8BE20 Cancer 1165 HE8BP05 Mixed Fetal 1166 HE8BP64 Cancer 1167 HE8BQ49 Mixed Fetal 1168 HE8BR18 Cancer 1169 HE8BR30 Mixed Fetal 1170 HE8BT58 Cancer 1171 HE8BU60 Cancer 1172 HE8CA13 Cancer 1173 HE8CC34 Cardiovascular, Digestive, Mixed Fetal 1174 HE8CH08 Cancer 1175 HE8DG02 Mixed Fetal 1176 HE8DK52 Cancer 1177 HE8DZ94 Cancer 1178 HE8EN79 Cancer 1179 HE8EX86 Cancer 1180 HE8FC10 Immune/Hematopoietic, Mixed Fetal, Reproductive 1181 HE8FG15 Cancer 1182 HE8FG24 Cancer 1183 HE8FK78 Cancer 1184 HE8FL24 Mixed Fetal 1185 HE8FL68 Mixed Fetal 1186 HE8FR53 Cancer 1187 HE8MA27 Cancer 1188 HE8MG56 Mixed Fetal 1189 HE8MQ01 Cancer 1190 HE8MS43 Cancer 1191 HE8MY77 Cancer 1192 HE8NC81 Cancer 1193 HE8NO09 Cancer 1194 HE8QU21 Immune/Hematopoietic, Mixed Fetal 1195 HE8SH74 Immune/Hematopoietic, Mixed Fetal, Musculoskeletal 1196 HE8UX34 Mixed Fetal 1197 HE9AE05 Cancer 1198 HE9BJ14 Mixed Fetal, Musculoskeletal 1199 HE9CI81 Cancer 1200 HE9CJ38 Mixed Fetal 1201 HE9CM11 Mixed Fetal 1202 HE9CN58 Cancer 1203 HE9CV59 Cancer 1204 HE9DG54 Cancer 1205 HE9DH59 Cancer 1206 HE9DZ47 Endocrine, Immune/Hematopoietic, Mixed Fetal 1207 HE9EC36 Cancer 1208 HE9EM54 Immune/Hematopoietic, Mixed Fetal 1209 HE9FH28 Mixed Fetal 1210 HE9HE13 Cancer 1211 HE9HE13 Cancer 1212 HE9HF59 Mixed Fetal 1213 HE9HV71 Cancer 1214 HE9NB82 Cancer 1215 HE9NE43 Mixed Fetal 1216 HE9RN58 Cancer 1217 HE9TA42 Cancer 1218 HEAAC21 Cancer 1219 HEAAC39 Neural/Sensory, Reproductive 1220 HEAAC48 Reproductive 1221 HEAAD63 Neural/Sensory, Reproductive 1222 HEAAE19 Immune/Hematopoietic, Reproductive 1223 HEAAM54 Reproductive 1224 HEAAM96 Reproductive 1225 HEAAN52 Cancer 1226 HEAAU28 Reproductive 1227 HEAAW54 Reproductive 1228 HEAAW94 Cancer 1229 HEBAP51 Cancer 1230 HEBAT05 Cancer 1231 HEBBF78 Cancer 1232 HEBBK04 Cancer 1233 HEBCN80 Neural/Sensory 1234 HEBCW57 Mixed Fetal, Neural/Sensory 1235 HEBDF90 Cancer 1236 HEBDW31 Cancer 1237 HEBFL36 Neural/Sensory 1238 HEBGC01 Neural/Sensory 1239 HEBGE23 Cancer 1240 HEBGE85 Digestive, Neural/Sensory, Reproductive 1241 HEBGJ94 Cancer 1242 HEBGM06 Cancer 1243 HEEAB58 Cancer 1244 HEEAF49 Cancer 1245 HEEAJ46 Mixed Fetal, Reproductive 1246 HEGAI20 Reproductive 1247 HEIAC52 Cancer 1248 HELAC55 Cardiovascular, Immune/Hematopoietic, Musculoskeletal 1249 HELAT58 Cardiovascular 1250 HELAW94 Cancer 1251 HELDF80 Cancer 1252 HELDH39 Cancer 1253 HELDK79 Cardiovascular 1254 HELDQ42 Cancer 1255 HELEE85 Cancer 1256 HELEL76 Cancer 1257 HELEL76 Cancer 1258 HELEO45 Cancer 1259 HELFA57 Cancer 1260 HELFO30 Cancer 1261 HELGF28 Cancer 1262 HELGP60 Cardiovascular, Excretory, Immune/Hematopoietic 1263 HELHN47 Cancer 1264 HELHP11 Cardiovascular, Immune/Hematopoietic 1265 HELHP11 Cardiovascular, Immune/Hematopoietic 1266 HEMAE30 Cancer 1267 HEMBV40 Cancer 1268 HEMCJ80 Cancer 1269 HEMCL55 Cardiovascular 1270 HEMDB07 Cardiovascular 1271 HEMDR05 Cardiovascular, Digestive, Immune/Hematopoietic 1272 HEMGK71 Cardiovascular, Immune/Hematopoietic, Musculoskeletal 1273 HEOMF59 Immune/Hematopoietic 1274 HEOMJ73 Cancer 1275 HEOMR67 Cancer 1276 HEOMU25 Connective/Epithelial, Immune/Hematopoietic 1277 HEOMU44 Cancer 1278 HEONI85 Digestive, Immune/Hematopoietic, Reproductive 1279 HEONK04 Cancer 1280 HEONP08 Immune/Hematopoietic 1281 HEPAD15 Endocrine, Reproductive 1282 HEPBC23 Cancer 1283 HEPBV09 Reproductive 1284 HEPCF35 Neural/Sensory, Reproductive 1285 HEPCU48 Cancer 1286 HEQAH47 Cancer 1287 HEQAP92 Cancer 1288 HEQAV53 Cancer 1289 HEQBJ01 Cancer 1290 HEQBJ01 Cancer 1291 HEQBJ01 Cancer 1292 HEQBM94 Cancer 1293 HEQCB93 Cancer 1294 HERAI63 Connective/Epithelial 1295 HERAQ22 Connective/Epithelial 1296 HERAS61 Connective/Epithelial 1297 HESAG57 Cancer 1298 HETAA62 Cancer 1299 HETBB70 Immune/Hematopoietic, Reproductive 1300 HETBJ88 Cancer 1301 HETCM67 Cancer 1302 HETDD61 Reproductive 1303 HETDD61 Reproductive 1304 HETDJ34 Cancer 1305 HETDM73 Cancer 1306 HETDP76 Cancer 1307 HETFO57 Cancer 1308 HETGZ31 Cancer 1309 HETHD26 Cancer 1310 HETHM27 Cancer 1311 HETIN36 Cancer 1312 HFAAI17 Neural/Sensory 1313 HFAAJ45 Immune/Hematopoietic, Neural/Sensory 1314 HFADF41 Neural/Sensory 1315 HFADM09 Cancer 1316 HFAUA23 Cancer 1317 HFCAG75 Cancer 1318 HFCAI40 Cancer 1319 HFCAQ17 Cancer 1320 HFCBC16 Neural/Sensory 1321 HFCBL53 Cancer 1322 HFCBL53 Cancer 1323 HFCBL53 Cancer 1324 HFCBT29 Cancer 1325 HFCCZ31 Cancer 1326 HFCDN13 Cancer 1327 HFCDT67 Cancer 1328 HFCDY36 Neural/Sensory 1329 HFCEC45 Cancer 1330 HFCET43 Cancer 1331 HFEAG55 Cancer 1332 HFEAU63 Connective/Epithelial 1333 HFEBA88 Cancer 1334 HFEBK75 Connective/Epithelial 1335 HFEBO15 Cancer 1336 HFEBO17 Cancer 1337 HFFAE46 Neural/Sensory 1338 HFFAH01 Digestive, Immune/Hematopoietic, Neural/Sensory 1339 HFFAL70 Cancer 1340 HFFAV61 Neural/Sensory 1341 HFGAB50 Cancer 1342 HFGAE28 Cancer 1343 HFGAN63 Cancer 1344 HFHDN80 Cardiovascular, Digestive, Immune/Hematopoietic 1345 HFIAB78 Cancer 1346 HFIAD23 Cancer 1347 HFIAK06 Musculoskeletal, Reproductive 1348 HFICH70 Musculoskeletal 1349 HFIHQ57 Musculoskeletal, Reproductive 1350 HFIIK29 Cancer 1351 HFIIK29 Cancer 1352 HFIIK29 Cancer 1353 HFIIK29 Cancer 1354 HFIIQ27 Cancer 1355 HFIIQ64 Cancer 1356 HFIIZ61 Cancer 1357 HFIJD81 Cancer 1358 HFIJF44 Cancer 1359 HFITA02 Immune/Hematopoietic, Musculoskeletal 1360 HFITF80 Cancer 1361 HFIUK66 Cancer 1362 HFIUT21 Cancer 1363 HFIVB04 Cancer 1364 HFIXC39 Cancer 1365 HFIXC69 Cancer 1366 HFIXE39 Cancer 1367 HFIYP15 Immune/Hematopoietic, Musculoskeletal 1368 HFIZE10 Cancer 1369 HFIZF51 Musculoskeletal 1370 HFIZK42 Immune/Hematopoietic, Musculoskeletal 1371 HFIZM89 Musculoskeletal 1372 HFKBA62 Digestive, Excretory, Neural/Sensory 1373 HFKBC47 Cancer 1374 HFKDX53 Cancer 1375 HFKEB14 Cancer 1376 HFKEG63 Excretory 1377 HFKES35 Cancer 1378 HFKES35 Cancer 1379 HFKEU17 Cancer 1380 HFKEV77 Cancer 1381 HFKFI15 Cancer 1382 HFKFI35 Excretory 1383 HFKFK49 Cancer 1384 HFKFV88 Cancer 1385 HFKFV88 Cancer 1386 HFKFV88 Cancer 1387 HFKFX64 Excretory 1388 HFOXD49 Cancer 1389 HFOXE28 Cancer 1390 HFOYH74 Musculoskeletal 1391 HFOYP02 Musculoskeletal 1392 HFOYR24 Musculoskeletal 1393 HFOYR54 Cancer 1394 HFOZB26 Cancer 1395 HFPBF54 Cancer 1396 HFPBF54 Cancer 1397 HFPBI93 Cancer 1398 HFPBJ64 Musculoskeletal, Neural/Sensory 1399 HFPBQ55 Musculoskeletal, Neural/Sensory, Reproductive 1400 HFPCK22 Cancer 1401 HFPCM32 Neural/Sensory 1402 HFPCM36 Cancer 1403 HFPCS84 Neural/Sensory 1404 HFPCU47 Neural/Sensory 1405 HFPCY66 Digestive, Neural/Sensory 1406 HFPDC65 Cancer 1407 HFPDE42 Musculoskeletal, Neural/Sensory 1408 HFPDE88 Neural/Sensory 1409 HFPDO25 Neural/Sensory 1410 HFPDP70 Neural/Sensory 1411 HFPDR39 Cancer 1412 HFPDX08 Cancer 1413 HFPEP69 Neural/Sensory 1414 HFRAU40 Cancer 1415 HFRAY90 Cancer 1416 HFSAY91 Cancer 1417 HFSBC10 Immune/Hematopoietic, Mixed Fetal 1418 HFSBE94 Immune/Hematopoietic 1419 HFTAN11 Cancer 1420 HFTAR27 Cancer 1421 HFTAR30 Cancer 1422 HFTAS49 Cancer 1423 HFTBB50 Cancer 1424 HFTBL17 Cancer 1425 HFTBL17 Cancer 1426 HFTCF02 Digestive, Musculoskeletal, Neural/Sensory 1427 HFTCI85 Neural/Sensory 1428 HFTCJ32 Neural/Sensory 1429 HFTCO17 Cancer 1430 HFTCW07 Neural/Sensory 1431 HFTDF32 Cancer 1432 HFTDF79 Immune/Hematopoietic, Neural/Sensory, Reproductive 1433 HFTDK11 Cancer 1434 HFTDU08 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 1435 HFVGK67 Digestive, Immune/Hematopoietic 1436 HFVHD38 Cancer 1437 HFVHY57 Cancer 1438 HFVIC33 Cancer 1439 HFXAK32 Cancer 1440 HFXAK59 Cancer 1441 HFXBI64 Neural/Sensory 1442 HFXBL05 Mixed Fetal, Neural/Sensory 1443 HFXBM52 Neural/Sensory 1444 HFXBR58 Neural/Sensory 1445 HFXBV67 Digestive, Neural/Sensory 1446 HFXBY20 Neural/Sensory 1447 HFXCB70 Neural/Sensory 1448 HFXCI42 Neural/Sensory 1449 HFXCL59 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 1450 HFXCM22 Neural/Sensory, Reproductive 1451 HFXCN18 Neural/Sensory 1452 HFXCS53 Cancer 1453 HFXDB37 Neural/Sensory 1454 HFXDI32 Neural/Sensory 1455 HFXDJ43 Neural/Sensory 1456 HFXDL76 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 1457 HFXDM75 Neural/Sensory 1458 HFXDO18 Neural/Sensory 1459 HFXDP44 Neural/Sensory 1460 HFXDR08 Immune/Hematopoietic, Neural/Sensory, Reproductive 1461 HFXDR28 Neural/Sensory 1462 HFXDR28 Neural/Sensory 1463 HFXDR47 Cancer 1464 HFXDZ03 Immune/Hematopoietic, Neural/Sensory 1465 HFXED33 Neural/Sensory 1466 HFXEE88 Neural/Sensory 1467 HFXGR32 Neural/Sensory 1468 HFXGT51 Neural/Sensory 1469 HFXGW16 Neural/Sensory 1470 HFXHC15 Neural/Sensory 1471 HFXHI33 Immune/Hematopoietic, Neural/Sensory 1472 HFXHL21 Neural/Sensory 1473 HFXHL83 Neural/Sensory 1474 HFXHM49 Neural/Sensory 1475 HFXHM93 Neural/Sensory 1476 HFXHN89 Immune/Hematopoietic, Neural/Sensory 1477 HFXJB21 Neural/Sensory 1478 HFXJN93 Neural/Sensory 1479 HFXJS15 Cancer 1480 HFXJT53 Cancer 1481 HFXKG56 Neural/Sensory 1482 HFXKL60 Cancer 1483 HFXLG08 Neural/Sensory 1484 HFXLK91 Cancer 1485 HFXLM32 Neural/Sensory, Reproductive 1486 HGBAX83 Digestive 1487 HGBBR29 Cancer 1488 HGBDL51 Cancer 1489 HGBDV35 Cancer 1490 HGBDX28 Cancer 1491 HGBGX31 Cancer 1492 HGBHE23 Cancer 1493 HGBHI15 Digestive 1494 HGCMW39 Cancer 1495 HGLAG32 Cancer 1496 HGLAH08 Cancer 1497 HGLAH86 Immune/Hematopoietic 1498 HGLBC33 Cancer 1499 HGLBG15 Cancer 1500 HGLBM55 Cancer 1501 HGLDA95 Cancer 1502 HGLDB06 Cancer 1503 HGLDE15 Digestive, Immune/Hematopoietic 1504 HHBEI14 Cancer 1505 HHBGL33 Cardiovascular, Digestive 1506 HHEAW44 Immune/Hematopoietic 1507 HHEBP28 Cancer 1508 HHECK41 Cancer 1509 HHECR10 Immune/Hematopoietic 1510 HHEMC55 Immune/Hematopoietic 1511 HHEMM20 Immune/Hematopoietic 1512 HHEMM80 Immune/Hematopoietic 1513 HHEMP35 Cancer 1514 HHEMZ08 Cancer 1515 HHENC17 Cancer 1516 HHENF95 Immune/Hematopoietic 1517 HHENR74 Immune/Hematopoietic 1518 HHENU33 Immune/Hematopoietic 1519 HHENY07 Cancer 1520 HHEOK77 Cancer 1521 HHEPE72 Immune/Hematopoietic 1522 HHEPE81 Cancer 1523 HHEPM64 Cancer 1524 HHEQI04 Connective/Epithelial, Excretory, Immune/Hematopoietic 1525 HHEQY60 Immune/Hematopoietic 1526 HHFBA31 Cancer 1527 HHFCI81 Cancer 1528 HHFCN78 Cardiovascular, Immune/Hematopoietic 1529 HHFCT95 Cancer 1530 HHFDN16 Cardiovascular 1531 HHFEB79 Connective/Epithelial 1532 HHFEC39 Cancer 1533 HHFEN34 Cardiovascular 1534 HHFFZ01 Cancer 1535 HHFGI71 Cardiovascular 1536 HHFGJ54 Cancer 1537 HHFGL38 Cardiovascular, Immune/Hematopoietic 1538 HHFGR75 Cardiovascular, Immune/Hematopoietic, Neural/Sensory 1539 HHFGZ23 Cardiovascular, Digestive, Endocrine 1540 HHFHG26 Cardiovascular, Neural/Sensory 1541 HHFHM47 Cardiovascular, Immune/Hematopoietic 1542 HHGAA76 Immune/Hematopoietic, Reproductive 1543 HHGAD46 Cancer 1544 HHGAT09 Cancer 1545 HHGBC21 Cancer 1546 HHGBF91 Cancer 1547 HHGBG63 Cancer 1548 HHGBV02 Immune/Hematopoietic, Reproductive 1549 HHGBW55 Immune/Hematopoietic, Reproductive 1550 HHGBX88 Cancer 1551 HHGCA26 Reproductive 1552 HHGDA81 Cancer 1553 HHGDI12 Neural/Sensory 1554 HHGDR05 Neural/Sensory 1555 HHGDR92 Cancer 1556 HHGDS56 Cancer 1557 HHGDW65 Cancer 1558 HHLBA86 Digestive 1559 HHNAC56 Digestive 1560 HHPBG90 Cancer 1561 HHPDE28 Cancer 1562 HHPDJ11 Cancer 1563 HHPDX86 Neural/Sensory 1564 HHPEA17 Neural/Sensory 1565 HHPEB61 Immune/Hematopoietic, Neural/Sensory, Respiratory 1566 HHPFP26 Cancer 1567 HHPFS11 Cardiovascular, Neural/Sensory 1568 HHPFS15 Cancer 1569 HHPFS18 Cancer 1570 HHPGH34 Neural/Sensory, Reproductive 1571 HHPGU74 Neural/Sensory 1572 HHPGU87 Cancer 1573 HHPSD42 Immune/Hematopoietic, Neural/Sensory 1574 HHPSE03 Neural/Sensory 1575 HHPSE55 Cancer 1576 HHPSF70 Cancer 1577 HHPSH74 Cancer 1578 HHPSL14 Cancer 1579 HHPSM40 Neural/Sensory 1580 HHPTF26 Mixed Fetal, Musculoskeletal, Neural/Sensory 1581 HHSAD31 Immune/Hematopoietic, Neural/Sensory, Reproductive 1582 HHSAE74 Neural/Sensory 1583 HHSAG62 Cancer 1584 HHSAK17 Neural/Sensory 1585 HHSBJ92 Cancer 1586 HHSBN84 Cancer 1587 HHSCL24 Cancer 1588 HHSCQ67 Cancer 1589 HHSCU12 Cancer 1590 HHSDB43 Cancer 1591 HHSDL07 Neural/Sensory 1592 HHSDX07 Cancer 1593 HHSFF54 Cancer 1594 HHSGB85 Cancer 1595 HHSGL84 Neural/Sensory 1596 HHTLH79 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 1597 HIABC70 Cancer 1598 HIATG10 Endocrine 1599 HIBCO70 Cancer 1600 HIBCR82 Mixed Fetal, Neural/Sensory 1601 HIBDA41 Cancer 1602 HIBEC45 Cancer 1603 HILBW03 Cancer 1604 HISAE16 Digestive 1605 HISAG53 Cancer 1606 HISAN63 Digestive 1607 HISAT78 Cancer 1608 HISBA38 Digestive, Immune/Hematopoietic 1609 HISBB66 Cancer 1610 HISCJ20 Cancer 1611 HISCK41 Digestive 1612 HISCO45 Cancer 1613 HISEJ52 Cancer 1614 HJABC58 Cancer 1615 HJABG59 Immune/Hematopoietic 1616 HJABR75 Immune/Hematopoietic 1617 HJABS31 Cancer 1618 HJABT12 Digestive, Immune/Hematopoietic, Neural/Sensory 1619 HJACE25 Cancer 1620 HJACK21 Cancer 1621 HJBCG74 Cancer 1622 HJBCO21 Cancer 1623 HJBCQ40 Immune/Hematopoietic 1624 HJBDM36 Cancer 1625 HJMAF30 Cancer 1626 HJMAM72 Cancer 1627 HJMAZ60 Cancer 1628 HJMBB20 Cancer 1629 HJMBB20 Cancer 1630 HJMBB20 Cancer 1631 HJMBK59 Cancer 1632 HJMBP01 Cancer 1633 HJMBQ17 Cancer 1634 HJMBW62 Reproductive 1635 HJMBX54 Musculoskeletal, Reproductive 1636 HJPAF69 Immune/Hematopoietic 1637 HJPAQ19 Cancer 1638 HJPAZ35 Cancer 1639 HJPBI77 Immune/Hematopoietic, Musculoskeletal, Neural/Sensory 1640 HJPBN96 Cancer 1641 HJPBU47 Cancer 1642 HJPCQ19 Cancer 1643 HJPDJ08 Immune/Hematopoietic 1644 HJPDK61 Cancer 1645 HKABI53 Cancer 1646 HKABN63 Cancer 1647 HKACA25 Cancer 1648 HKACO64 Cancer 1649 HKACP50 Cancer 1650 HKACX90 Cancer 1651 HKADI27 Cancer 1652 HKADN26 Cancer 1653 HKADP79 Cancer 1654 HKADT55 Cancer 1655 HKAEK58 Cancer 1656 HKAEK72 Connective/Epithelial 1657 HKAFJ47 Cancer 1658 HKAFQ41 Cancer 1659 HKAHH71 Cancer 1660 HKAJA95 Cancer 1661 HKAKU90 Cancer 1662 HKCSZ54 Digestive 1663 HKFAA15 Cancer 1664 HKFBB08 Immune/Hematopoietic 1665 HKGAG59 Cancer 1666 HKGAJ81 Cancer 1667 HKGAK45 Musculoskeletal, Reproductive 1668 HKGAP04 Cancer 1669 HKGAP57 Immune/Hematopoietic 1670 HKGAW41 Cancer 1671 HKGBA21 Connective/Epithelial, Mixed Fetal, Musculoskeletal 1672 HKGBC33 Immune/Hematopoietic 1673 HKGBC73 Cancer 1674 HKGBF61 Cancer 1675 HKGBH54 Cancer 1676 HKGBP52 Cancer 1677 HKGCE23 Cancer 1678 HKGCE62 Immune/Hematopoietic 1679 HKGCK41 Cancer 1680 HKGCK41 Cancer 1681 HKGCN96 Cancer 1682 HKGCX05 Cancer 1683 HKGDA95 Cancer 1684 HKGDO12 Cancer 1685 HKIME53 Cancer 1686 HKIMG23 Cancer 1687 HKIXB73 Excretory 1688 HKIXD68 Cancer 1689 HKIXR91 Cancer 1690 HKIXS19 Cancer 1691 HKIXW45 Cancer 1692 HKIYU90 Excretory, Neural/Sensory 1693 HKMLB81 Excretory 1694 HKMLF77 Excretory 1695 HKMLM32 Excretory, Neural/Sensory 1696 HKMLR17 Cancer 1697 HKMLT89 Excretory, Immune/Hematopoietic, Reproductive 1698 HKMLV05 Excretory, Immune/Hematopoietic 1699 HKMLV25 Cancer 1700 HKMMB79 Cancer 1701 HKMMC69 Excretory, Immune/Hematopoietic 1702 HKMMD91 Cancer 1703 HKMMP90 Excretory, Immune/Hematopoietic Neural/Sensory 1704 HKMMU76 Cancer 1705 HKPAC10 Excretory 1706 HKPAC50 Cancer 1707 HKPMA08 Cancer 1708 HKTAC18 Cancer 1709 HL1SA89 Cancer 1710 HL2AB60 Cancer 1711 HL3AE69 Cancer 1712 HL3AF32 Cancer 1713 HLDAV70 Digestive, Immune/Hematopoietic 1714 HLDBL62 Cancer 1715 HLDBV18 Cancer 1716 HLDBV54 Cancer 1717 HLDCR26 Cancer 1718 HLDDM27 Cancer 1719 HLDDM27 Cancer 1720 HLDNF18 Cancer 1721 HLDNN84 Digestive, Mixed Fetal 1722 HLDOD77 Digestive 1723 HLDOL74 Cancer 1724 HLDPB24 Cardiovascular, Digestive 1725 HLDRU08 Cancer 1726 HLDXF43 Cancer 1727 HLEAA10 Immune/Hematopoietic 1728 HLEAA24 Immune/Hematopoietic 1729 HLHAE14 Neural/Sensory, Respiratory 1730 HLHAE14 Neural/Sensory, Respiratory 1731 HLHBS54 Cancer 1732 HLHCB33 Digestive, Reproductive, Respiratory 1733 HLHCF14 Connective/Epithelial, Respiratory 1734 HLHCG24 Cancer 1735 HLHCH20 Cancer 1736 HLHCN51 Digestive, Immune/Hematopoietic, Respiratory 1737 HLHCT96 Cancer 1738 HLHDC33 Immune/Hematopoietic, Reproductive, Respiratory 1739 HLHDF92 Cancer 1740 HLHDJ05 Respiratory 1741 HLHDL37 Respiratory 1742 HLHDL69 Cancer 1743 HLHDL69 Cancer 1744 HLHDL69 Cancer 1745 HLHDL69 Cancer 1746 HLHDM38 Cancer 1747 HLHDR92 Neural/Sensory, Respiratory 1748 HLHDY94 Cancer 1749 HLHEE27 Cancer 1750 HLHEE38 Connective/Epithelial, Neural/Sensory, Respiratory 1751 HLHEI72 Musculoskeletal, Respiratory 1752 HLHEX62 Excretory, Immune/Hematopoietic, Respiratory 1753 HLHFK59 Digestive, Respiratory 1754 HLHFP09 Cancer 1755 HLHGG78 Cancer 1756 HLHSG15 Cancer 1757 HLHSQ35 Respiratory 1758 HLHTB92 Immune/Hematopoietic, Respiratory 1759 HLHTP55 Cancer 1760 HLIBD74 Digestive 1761 HLIBE41 Digestive, Immune/Hematopoietic, Reproductive 1762 HLIBO16 Digestive, Immune/Hematopoietic 1763 HLJBI22 Cancer 1764 HLJEE16 Cancer 1765 HLLAX64 Cancer 1766 HLLAX95 Immune/Hematopoietic 1767 HLLCD67 Immune/Hematopoietic 1768 HLMBX89 Cancer 1769 HLMBZ14 Immune/Hematopoietic 1770 HLMCT51 Immune/Hematopoietic, Reproductive 1771 HLMCT95 Cancer 1772 HLMDD65 Cancer 1773 HLMDH01 Immune/Hematopoietic 1774 HLMDU23 Immune/Hematopoietic 1775 HLMFB62 Immune/Hematopoietic 1776 HLMFG52 Immune/Hematopoietic 1777 HLMFU53 Cancer 1778 HLMHG68 Cancer 1779 HLMHN06 Immune/Hematopoietic, Neural/Sensory 1780 HLMHS15 Immune/Hematopoietic 1781 HLMIM84 Cancer 1782 HLMIN52 Cancer 1783 HLMIQ83 Immune/Hematopoietic 1784 HLMIW76 Immune/Hematopoietic 1785 HLMMA65 Immune/Hematopoietic 1786 HLMMT12 Digestive, Immune/Hematopoietic 1787 HLMNA19 Cardiovascular, Immune/Hematopoietic 1788 HLQAD72 Cancer 1789 HLQAM30 Cancer 1790 HLQAM59 Digestive 1791 HLQBB23 Cancer 1792 HLQBF05 Digestive, Reproductive 1793 HLQBX64 Cancer 1794 HLQCY09 Digestive 1795 HLQCZ43 Cancer 1796 HLQCZ80 Digestive 1797 HLQDK45 Digestive 1798 HLQDM47 Digestive 1799 HLQDU77 Cancer 1800 HLSAD72 Connective/Epithelial 1801 HLTCJ67 Immune/Hematopoietic 1802 HLTCM28 Immune/Hematopoietic, Respiratory 1803 HLTCO22 Cancer 1804 HLTDA14 Immune/Hematopoietic 1805 HLTDC26 Cancer 1806 HLTDC26 Cancer 1807 HLTDI20 Cancer 1808 HLTDI65 Immune/Hematopoietic 1809 HLTDK30 Cancer 1810 HLTDL37 Cancer 1811 HLTDU35 Cancer 1812 HLTDX04 Cancer 1813 HLTEH84 Cancer 1814 HLTEL39 Cardiovascular, Immune/Hematopoietic 1815 HLTEN11 Cancer 1816 HLTEW52 Immune/Hematopoietic 1817 HLTEZ36 Cancer 1818 HLTGG14 Cancer 1819 HLUAF94 Immune/Hematopoietic 1820 HLWAH33 Mixed Fetal, Reproductive 1821 HLWAO11 Cancer 1822 HLWAW73 Cancer 1823 HLWAX50 Cancer 1824 HLWBJ93 Cancer 1825 HLWBK16 Cardiovascular, Connective/Epithelial, Reproductive 1826 HLWCC11 Reproductive 1827 HLYAH81 Immune/Hematopoietic 1828 HLYAH92 Immune/Hematopoietic 1829 HLYAJ79 Cancer 1830 HLYAL28 Immune/Hematopoietic 1831 HLYAR30 Cancer 1832 HLYAT54 Immune/Hematopoietic 1833 HLYBC81 Connective/Epithelial, Immune/Hematopoietic, Reproductive 1834 HLYBD09 Immune/Hematopoietic 1835 HLYBL67 Immune/Hematopoietic 1836 HLYBM38 Digestive, Immune/Hematopoietic 1837 HLYBN23 Immune/Hematopoietic 1838 HLYBN71 Immune/Hematopoietic 1839 HLYBS25 Digestive, Immune/Hematopoietic, Reproductive 1840 HLYBT28 Immune/Hematopoietic 1841 HLYBU15 Immune/Hematopoietic 1842 HLYBY04 Immune/Hematopoietic 1843 HLYCE15 Digestive, Immune/Hematopoietic 1844 HLYCH04 Immune/Hematopoietic 1845 HLYCY48 Immune/Hematopoietic 1846 HLYDE38 Immune/Hematopoietic 1847 HLYDG55 Immune/Hematopoietic 1848 HLYDO73 Immune/Hematopoietic 1849 HLYEA60 Cancer 1850 HLYEJ14 Cancer 1851 HLYEJ44 Cancer 1852 HLYEU51 Immune/Hematopoietic 1853 HLYGV19 Cancer 1854 HMABK52 Immune/Hematopoietic 1855 HMACF34 Immune/Hematopoietic 1856 HMACL77 Cancer 1857 HMACT74 Immune/Hematopoietic 1858 HMADJ14 Connective/Epithelial, Immune/Hematopoietic, Musculoskeletal 1859 HMADJ74 Connective/Epithelial, Immune/Hematopoietic, Musculoskeletal 1860 HMAEA58 Cancer 1861 HMAGF01 Cancer 1862 HMAJS26 Cancer 1863 HMCED78 Cancer 1864 HMCFN86 Cancer 1865 HMCGJ47 Cancer 1866 HMCGK88 Cancer 1867 HMCIH27 Cancer 1868 HMCIQ20 Cardiovascular, Immune/Hematopoietic, Neural/Sensory 1869 HMCJC19 Immune/Hematopoietic 1870 HMDAB44 Neural/Sensory 1871 HMDAE88 Neural/Sensory 1872 HMDAG62 Cancer 1873 HMDAK20 Neural/Sensory 1874 HMDAM08 Neural/Sensory 1875 HMDAM39 Neural/Sensory 1876 HMEAA41 Cancer 1877 HMECM77 Cardiovascular 1878 HMEEH21 Cardiovascular 1879 HMEET36 Cancer 1880 HMEEZ07 Cardiovascular, Reproductive 1881 HMEFB15 Cardiovascular 1882 HMEIH57 Cardiovascular, Immune/Hematopoietic 1883 HMEIJ21 Cancer 1884 HMEIX79 Cancer 1885 HMEJC96 Cancer 1886 HMEJD36 Cardiovascular, Endocrine, Immune/Hematopoietic 1887 HMEJK28 Cancer 1888 HMEKH55 Cancer 1889 HMEKW44 Cardiovascular, Immune/Hematopoietic, Neural/Sensory 1890 HMEKW71 Cancer 1891 HMELW26 Cancer 1892 HMGBT32 Cancer 1893 HMHBI09 Cancer 1894 HMHBI93 Cancer 1895 HMHBP74 Cancer 1896 HMIAC52 Cancer 1897 HMIAD75 Neural/Sensory 1898 HMIAG42 Cancer 1899 HMIAG55 Cancer 1900 HMIAG72 Cancer 1901 HMIAL39 Cancer 1902 HMIAO82 Cancer 1903 HMIAR42 Cancer 1904 HMIAV33 Immune/Hematopoietic, Mixed Fetal, Neural/Sensory 1905 HMIAZ24 Cancer 1906 HMIBD93 Cancer 1907 HMIBE95 Neural/Sensory 1908 HMIBG57 Immune/Hematopoietic, Neural/Sensory, Reproductive 1909 HMJAC12 Neural/Sensory 1910 HMKAN71 Cancer 1911 HMKBA33 Neural/Sensory 1912 HMKCI22 Cancer 1913 HMKCK32 Neural/Sensory 1914 HMKCP81 Cancer 1915 HMKCY49 Immune/Hematopoietic, Neural/Sensory 1916 HMKDD51 Neural/Sensory 1917 HMKDG69 Cardiovascular, Neural/Sensory 1918 HMKDM80 Neural/Sensory 1919 HMKEG88 Neural/Sensory 1920 HMMAA09 Immune/Hematopoietic 1921 HMMAK92 Immune/Hematopoietic 1922 HMMAL32 Immune/Hematopoietic 1923 HMMBD19 Immune/Hematopoietic, Reproductive 1924 HMMBF22 Immune/Hematopoietic, Reproductive 1925 HMMBH91 Immune/Hematopoietic 1926 HMMBH94 Immune/Hematopoietic 1927 HMMBK55 Immune/Hematopoietic 1928 HMMBQ31 Cardiovascular, Immune/Hematopoietic 1929 HMMBR63 Cancer 1930 HMMBS55 Immune/Hematopoietic, Reproductive 1931 HMMBT47 Immune/Hematopoietic 1932 HMMCD35 Immune/Hematopoietic 1933 HMMCD95 Immune/Hematopoietic, Neural/Sensory 1934 HMPAB26 Cancer 1935 HMPAP48 Immune/Hematopoietic 1936 HMQAI38 Immune/Hematopoietic, Reproductive 1937 HMQAT69 Cancer 1938 HMQBL90 Digestive, Immune/Hematopoietic 1939 HMQBV82 Immune/Hematopoietic, Musculoskeletal, Reproductive 1940 HMQCA75 Cancer 1941 HMQCB37 Cancer 1942 HMQCL80 Immune/Hematopoietic 1943 HMQCX41 Immune/Hematopoietic 1944 HMQDM09 Cancer 1945 HMQDU07 Digestive, Immune/Hematopoietic, Musculoskeletal 1946 HMSAP33 Immune/Hematopoietic 1947 HMSAZ48 Immune/Hematopoietic 1948 HMSBN18 Cancer 1949 HMSBS25 Immune/Hematopoietic 1950 HMSBU14 Immune/Hematopoietic 1951 HMSBZ10 Immune/Hematopoietic 1952 HMSCB94 Immune/Hematopoietic, Reproductive 1953 HMSCK12 Immune/Hematopoietic 1954 HMSCP63 Immune/Hematopoietic 1955 HMSCV75 Immune/Hematopoietic 1956 HMSCV85 Immune/Hematopoietic, Reproductive 1957 HMSCW44 Immune/Hematopoietic, Mixed Fetal, Neural/Sensory 1958 HMSCZ19 Cancer 1959 HMSDI67 Digestive, Immune/Hematopoietic 1960 HMSDI79 Cancer 1961 HMSDR28 Cancer 1962 HMSFT25 Immune/Hematopoietic 1963 HMSFW52 Immune/Hematopoietic 1964 HMSGT73 Immune/Hematopoietic 1965 HMSGU30 Cancer 1966 HMSHB42 Cancer 1967 HMSHB42 Cancer 1968 HMSHN72 Immune/Hematopoietic, Reproductive 1969 HMSHT29 Immune/Hematopoietic 1970 HMSHW73 Immune/Hematopoietic 1971 HMSIC48 Cardiovascular, Immune/Hematopoietic 1972 HMSII36 Immune/Hematopoietic 1973 HMSIT42 Digestive, Immune/Hematopoietic, Neural/Sensory 1974 HMSJB08 Cancer 1975 HMSJI69 Immune/Hematopoietic 1976 HMSJM20 Immune/Hematopoietic 1977 HMSJR44 Immune/Hematopoietic 1978 HMSKQ91 Immune/Hematopoietic 1979 HMSKY45 Immune/Hematopoietic 1980 HMTAF92 Cancer 1981 HMTAT36 Cancer 1982 HMUAB93 Cancer 1983 HMUAD65 Immune/Hematopoietic, Musculoskeletal 1984 HMUAT23 Cancer 1985 HMUBA47 Cancer 1986 HMUBJ22 Cancer 1987 HMUBK53 Cancer 1988 HMUBN24 Musculoskeletal 1989 HMUBO15 Cancer 1990 HMUBX48 Musculoskeletal, Reproductive 1991 HMUBY57 Cancer 1992 HMUBZ15 Cancer 1993 HMVAL15 Cancer 1994 HMVBC84 Digestive, Immune/Hematopoietic, Neural/Sensory 1995 HMVBD68 Cancer 1996 HMVCG17 Immune/Hematopoietic 1997 HMVCS92 Cancer 1998 HMVCS92 Cancer 1999 HMVDB45 Immune/Hematopoietic 2000 HMVDJ71 Cancer 2001 HMVDT89 Cancer 2002 HMVDT89 Cancer 2003 HMWAO65 Cancer 2004 HMWAO82 Immune/Hematopoietic 2005 HMWBD74 Cancer 2006 HMWBK35 Cancer 2007 HMWBK86 Immune/Hematopoietic, Mixed Fetal 2008 HMWBL38 Connective/Epithelial, Immune/Hematopoietic 2009 HMWBM48 Cancer 2010 HMWCG28 Cancer 2011 HMWCP85 Digestive, Immune/Hematopoietic 2012 HMWDG30 Cancer 2013 HMWDU20 Cancer 2014 HMWDX57 Digestive, Immune/Hematopoietic, Respiratory 2015 HMWDZ63 Immune/Hematopoietic 2016 HMWEA77 Immune/Hematopoietic 2017 HMWEC03 Cancer 2018 HMWEF46 Immune/Hematopoietic 2019 HMWEK43 Immune/Hematopoietic 2020 HMWEM23 Cancer 2021 HMWEM23 Cancer 2022 HMWER46 Cancer 2023 HMWEU96 Cancer 2024 HMWEX02 Cancer 2025 HMWFB65 Cancer 2026 HMWFD77 Immune/Hematopoietic 2027 HMWFO25 Immune/Hematopoietic 2028 HMWFO89 Cancer 2029 HMWGM41 Cancer 2030 HMWGO95 Immune/Hematopoietic 2031 HMWGV85 Cancer 2032 HMWGZ42 Immune/Hematopoietic 2033 HMWHR36 Immune/Hematopoietic 2034 HMWIM55 Immune/Hematopoietic 2035 HMWIQ26 Cancer 2036 HMWIU49 Cancer 2037 HMWJJ62 Cancer 2038 HMWJJ64 Cancer 2039 HNAAD76 Digestive, Immune/Hematopoietic 2040 HNAAE24 Digestive 2041 HNALD94 Cancer 2042 HNALE44 Cancer 2043 HNDAC35 Cancer 2044 HNEAA04 Immune/Hematopoietic 2045 HNEAH26 Immune/Hematopoietic, Neural/Sensory 2046 HNEAK38 Immune/Hematopoietic 2047 HNEAK65 Cancer 2048 HNEBX72 Immune/Hematopoietic, Neural/Sensory 2049 HNEBY79 Immune/Hematopoietic 2050 HNECD52 Immune/Hematopoietic, Neural/Sensory 2051 HNECL75 Cancer 2052 HNECX90 Cancer 2053 HNECX90 Cancer 2054 HNEDA05 Immune/Hematopoietic 2055 HNEDP75 Immune/Hematopoietic 2056 HNEDQ02 Cancer 2057 HNEDU46 Cancer 2058 HNFAD50 Cancer 2059 HNFAD50 Cancer 2060 HNFAG67 Cancer 2061 HNFCJ77 Immune/Hematopoietic, Reproductive 2062 HNFCO56 Cancer 2063 HNFCY57 Connective/Epithelial, Immune/Hematopoietic, Respiratory 2064 HNFDL89 Digestive, Immune/Hematopoietic 2065 HNFDT73 Excretory, Immune/Hematopoietic, Reproductive 2066 HNFDU92 Immune/Hematopoietic 2067 HNFDY09 Digestive, Immune/Hematopoietic, Neural/Sensory 2068 HNFDY31 Cancer 2069 HNFEA17 Cancer 2070 HNFEP55 Cancer 2071 HNFET12 Immune/Hematopoietic 2072 HNFFR59 Immune/Hematopoietic 2073 HNFGC51 Cancer 2074 HNFGR15 Immune/Hematopoietic 2075 HNFGW37 Immune/Hematopoietic 2076 HNFGW53 Cancer 2077 HNFHA34 Cancer 2078 HNFHD58 Cancer 2079 HNFHV68 Immune/Hematopoietic 2080 HNFIE15 Cancer 2081 HNFIE29 Immune/Hematopoietic 2082 HNFIG49 Immune/Hematopoietic 2083 HNFJE27 Immune/Hematopoietic 2084 HNFJG16 Immune/Hematopoietic 2085 HNGAC71 Digestive, Immune/Hematopoietic 2086 HNGAK42 Immune/Hematopoietic 2087 HNGAL25 Immune/Hematopoietic 2088 HNGAT83 Immune/Hematopoietic 2089 HNGAX06 Cancer 2090 HNGBB09 Immune/Hematopoietic 2091 HNGBC53 Immune/Hematopoietic 2092 HNGBD94 Immune/Hematopoietic 2093 HNGBE44 Digestive, Immune/Hematopoietic 2094 HNGBE63 Immune/Hematopoietic 2095 HNGBI83 Immune/Hematopoietic 2096 HNGBJ74 Immune/Hematopoietic 2097 HNGBP30 Immune/Hematopoietic 2098 HNGBQ61 Immune/Hematopoietic 2099 HNGBS35 Immune/Hematopoietic 2100 HNGBW25 Immune/Hematopoietic 2101 HNGCF29 Immune/Hematopoietic 2102 HNGCF64 Immune/Hematopoietic 2103 HNGDF54 Cancer 2104 HNGDH22 Immune/Hematopoietic 2105 HNGDH27 Immune/Hematopoietic 2106 HNGDN07 Immune/Hematopoietic, Reproductive 2107 HNGDO65 Cancer 2108 HNGDR39 Immune/Hematopoietic 2109 HNGDW78 Immune/Hematopoietic 2110 HNGEA90 Immune/Hematopoietic 2111 HNGEC17 Immune/Hematopoietic 2112 HNGEE06 Immune/Hematopoietic 2113 HNGEF70 Immune/Hematopoietic 2114 HNGEF72 Immune/Hematopoietic 2115 HNGEI64 Immune/Hematopoietic 2116 HNGEJ33 Cancer 2117 HNGEK64 Immune/Hematopoietic 2118 HNGEN32 Immune/Hematopoietic 2119 HNGER85 Immune/Hematopoietic 2120 HNGES90 Immune/Hematopoietic 2121 HNGET33 Immune/Hematopoietic 2122 HNGEX18 Immune/Hematopoietic 2123 HNGEY45 Immune/Hematopoietic 2124 HNGEZ02 Immune/Hematopoietic, Reproductive 2125 HNGEZ90 Immune/Hematopoietic 2126 HNGFA25 Immune/Hematopoietic 2127 HNGFB05 Immune/Hematopoietic 2128 HNGFD30 Immune/Hematopoietic, Mixed Fetal 2129 HNGFD31 Immune/Hematopoietic 2130 HNGFD61 Excretory, Immune/Hematopoietic 2131 HNGFG04 Immune/Hematopoietic 2132 HNGFG74 Immune/Hematopoietic 2133 HNGFH32 Immune/Hematopoietic 2134 HNGFH83 Immune/Hematopoietic 2135 HNGFI21 Cancer 2136 HNGFM31 Immune/Hematopoietic 2137 HNGFN77 Immune/Hematopoietic 2138 HNGFQ18 Immune/Hematopoietic 2139 HNGFR54 Immune/Hematopoietic 2140 HNGFT70 Immune/Hematopoietic 2141 HNGFU70 Immune/Hematopoietic 2142 HNGFV39 Immune/Hematopoietic 2143 HNGGF13 Immune/Hematopoietic 2144 HNGGK63 Immune/Hematopoietic 2145 HNGGK65 Immune/Hematopoietic, Neural/Sensory, Reproductive 2146 HNGGL11 Immune/Hematopoietic 2147 HNGGO05 Immune/Hematopoietic 2148 HNGGS92 Immune/Hematopoietic 2149 HNGGT10 Cancer 2150 HNGGT74 Immune/Hematopoietic 2151 HNGHB89 Digestive, Immune/Hematopoietic, Reproductive 2152 HNGHD07 Immune/Hematopoietic 2153 HNGHK37 Immune/Hematopoietic 2154 HNGHM47 Immune/Hematopoietic 2155 HNGHT01 Immune/Hematopoietic, Neural/Sensory 2156 HNGHT86 Immune/Hematopoietic 2157 HNGIH40 Immune/Hematopoietic 2158 HNGIK07 Connective/Epithelial, Immune/Hematopoietic, Musculoskeletal 2159 HNGIM40 Immune/Hematopoietic 2160 HNGIM83 Immune/Hematopoietic 2161 HNGIO93 Cancer 2162 HNGIS27 Immune/Hematopoietic 2163 HNGIU16 Immune/Hematopoietic, Reproductive 2164 HNGIX91 Immune/Hematopoietic 2165 HNGJA68 Immune/Hematopoietic 2166 HNGJB57 Immune/Hematopoietic 2167 HNGJE86 Immune/Hematopoietic 2168 HNGJH26 Immune/Hematopoietic 2169 HNGJJ61 Immune/Hematopoietic 2170 HNGJL07 Immune/Hematopoietic, Neural/Sensory 2171 HNGJS66 Immune/Hematopoietic, Reproductive 2172 HNGJU60 Immune/Hematopoietic 2173 HNGKB09 Immune/Hematopoietic, Reproductive 2174 HNGKW35 Immune/Hematopoietic 2175 HNGKY94 Immune/Hematopoietic 2176 HNGLD28 Immune/Hematopoietic 2177 HNGOY36 Immune/Hematopoietic 2178 HNHAB38 Immune/Hematopoietic, Musculoskeletal 2179 HNHAC43 Cancer 2180 HNHAD34 Immune/Hematopoietic 2181 HNHAG83 Immune/Hematopoietic, Mixed Fetal, Musculoskeletal 2182 HNHAH06 Immune/Hematopoietic 2183 HNHAJ65 Immune/Hematopoietic 2184 HNHAL61 Immune/Hematopoietic 2185 HNHAP58 Cancer 2186 HNHAW34 Immune/Hematopoietic 2187 HNHAW35 Immune/Hematopoietic 2188 HNHAY26 Immune/Hematopoietic 2189 HNHAY86 Immune/Hematopoietic, Neural/Sensory 2190 HNHAZ20 Immune/Hematopoietic 2191 HNHBE21 Immune/Hematopoietic 2192 HNHBE38 Cancer 2193 HNHBG18 Immune/Hematopoietic 2194 HNHBI65 Immune/Hematopoietic 2195 HNHBM16 Immune/Hematopoietic, Neural/Sensory 2196 HNHCH78 Immune/Hematopoietic 2197 HNHCP14 Immune/Hematopoietic 2198 HNHCQ44 Immune/Hematopoietic 2199 HNHCT22 Cardiovascular, Immune/Hematopoietic 2200 HNHCT47 Cardiovascular, Immune/Hematopoietic 2201 HNHCV48 Immune/Hematopoietic 2202 HNHCZ54 Cancer 2203 HNHDC52 Immune/Hematopoietic, Neural/Sensory 2204 HNHDD95 Immune/Hematopoietic 2205 HNHDE58 Cancer 2206 HNHDI17 Immune/Hematopoietic 2207 HNHDL37 Immune/Hematopoietic 2208 HNHDM21 Immune/Hematopoietic 2209 HNHDR57 Immune/Hematopoietic 2210 HNHDR96 Immune/Hematopoietic 2211 HNHDU62 Immune/Hematopoietic 2212 HNHDW34 Immune/Hematopoietic 2213 HNHDX28 Immune/Hematopoietic 2214 HNHDZ06 Immune/Hematopoietic 2215 HNHDZ42 Immune/Hematopoietic 2216 HNHEF37 Immune/Hematopoietic 2217 HNHEF49 Immune/Hematopoietic 2218 HNHEF70 Immune/Hematopoietic 2219 HNHEG30 Immune/Hematopoietic 2220 HNHEH38 Immune/Hematopoietic 2221 HNHEL22 Immune/Hematopoietic 2222 HNHEN70 Cancer 2223 HNHEP21 Immune/Hematopoietic 2224 HNHEP41 Immune/Hematopoietic 2225 HNHES33 Immune/Hematopoietic, Reproductive 2226 HNHET16 Immune/Hematopoietic 2227 HNHEY29 Immune/Hematopoietic 2228 HNHEZ76 Immune/Hematopoietic 2229 HNHFF60 Immune/Hematopoietic 2230 HNHFF81 Immune/Hematopoietic, Neural/Sensory 2231 HNHFJ49 Immune/Hematopoietic 2232 HNHFR42 Immune/Hematopoietic 2233 HNHFX25 Immune/Hematopoietic, Musculoskeletal 2234 HNHGD95 Cardiovascular, Immune/Hematopoietic 2235 HNHGR82 Immune/Hematopoietic 2236 HNHGS62 Immune/Hematopoietic 2237 HNHGY77 Cancer 2238 HNHHA47 Digestive, Immune/Hematopoietic, Respiratory 2239 HNHHN22 Immune/Hematopoietic 2240 HNHHW53 Immune/Hematopoietic 2241 HNHIB40 Immune/Hematopoietic 2242 HNHKI74 Immune/Hematopoietic 2243 HNHKV56 Immune/Hematopoietic 2244 HNHLD80 Immune/Hematopoietic 2245 HNHLS76 Immune/Hematopoietic 2246 HNHLZ47 Immune/Hematopoietic 2247 HNHMP15 Immune/Hematopoietic 2248 HNHMP62 Immune/Hematopoietic 2249 HNHMY76 Immune/Hematopoietic, Reproductive 2250 HNHMZ01 Immune/Hematopoietic 2251 HNHND14 Immune/Hematopoietic 2252 HNHND94 Immune/Hematopoietic 2253 HNHOF09 Immune/Hematopoietic 2254 HNKAA76 Cancer 2255 HNTAF42 Cancer 2256 HNTCG32 Cancer 2257 HNTNY89 Cancer 2258 HNTRB25 Cancer 2259 HNTRQ40 Cancer 2260 HNTSQ23 Cancer 2261 HOAAH51 Cancer 2262 HOAAI76 Cancer 2263 HOAAJ09 Cancer 2264 HOAAL10 Musculoskeletal 2265 HOAAU13 Mixed Fetal, Musculoskeletal 2266 HOABC12 Cancer 2267 HOABH36 Cancer 2268 HOBNA89 Musculoskeletal 2269 HOBNF51 Cancer 2270 HODAH24 Reproductive 2271 HODAH46 Cancer 2272 HODAV25 Cancer 2273 HODAW64 Cardiovascular, Neural/Sensory, Reproductive 2274 HODAY17 Cancer 2275 HODBA45 Reproductive 2276 HODBC79 Cancer 2277 HODBD79 Immune/Hematopoietic, Reproductive 2278 HODBF12 Cancer 2279 HODBF86 Digestive, Reproductive 2280 HODBF91 Cancer 2281 HODBW34 Digestive, Immune/Hematopoietic, Reproductive 2282 HODBX93 Reproductive 2283 HODBZ06 Cancer 2284 HODCA73 Cancer 2285 HODCV86 Immune/Hematopoietic, Reproductive 2286 HODCY44 Reproductive 2287 HODDB58 Neural/Sensory, Reproductive 2288 HODDG72 Cancer 2289 HODDJ25 Cancer 2290 HODDN21 Reproductive 2291 HODDO31 Reproductive 2292 HODDQ06 Cancer 2293 HODEA20 Cancer 2294 HODEM38 Digestive, Immune/Hematopoietic, Reproductive 2295 HODET37 Reproductive 2296 HOEBI94 Cancer 2297 HOEBJ70 Cancer 2298 HOECB33 Cancer 2299 HOECX21 Cancer 2300 HOEDE27 Musculoskeletal 2301 HOEEK81 Cancer 2302 HOEEZ62 Musculoskeletal 2303 HOEFJ26 Cancer 2304 HOEFL74 Cardiovascular, Digestive, Musculoskeletal 2305 HOFMF63 Cancer 2306 HOFMJ65 Cancer 2307 HOFMK02 Immune/Hematopoietic, Reproductive 2308 HOFMO16 Reproductive 2309 HOFMP62 Reproductive 2310 HOFMT59 Reproductive 2311 HOFMV22 Reproductive 2312 HOFND06 Digestive, Reproductive 2313 HOFNY15 Reproductive 2314 HOFNY28 Reproductive 2315 HOFOC41 Reproductive 2316 HOGAA41 Cancer 2317 HOGAB51 Immune/Hematopoietic, Reproductive 2318 HOGAH40 Cancer 2319 HOGAP06 Immune/Hematopoietic, Reproductive 2320 HOGAR36 Reproductive 2321 HOGAR71 Cancer 2322 HOGCC26 Cancer 2323 HOGCD78 Reproductive 2324 HOGCK03 Cancer 2325 HOGCL01 Cancer 2326 HOHBB36 Cancer 2327 HOHBC57 Cancer 2328 HOHBO66 Cancer 2329 HOHBZ10 Cancer 2330 HOHCH71 Cancer 2331 HOHEB48 Musculoskeletal 2332 HONAH67 Digestive, Excretory, Reproductive 2333 HOOAC84 Immune/Hematopoietic, Neural/Sensory, Reproductive 2334 HOPBP13 Cancer 2335 HOQBG21 Cancer 2336 HORBI80 Cancer 2337 HORBL77 Cancer 2338 HOSBX14 Immune/Hematopoietic, Musculoskeletal, Reproductive 2339 HOSCZ41 Cancer 2340 HOSEM81 Cancer 2341 HOSEO83 Cancer 2342 HOSFR35 Cancer 2343 HOUAZ32 Cancer 2344 HOUBC29 Connective/Epithelial, Immune/Hematopoietic, Reproductive 2345 HOUBG39 Connective/Epithelial, Immune/Hematopoietic, Musculoskeletal 2346 HOUCD12 Connective/Epithelial 2347 HOUDB17 Connective/Epithelial 2348 HOUDX40 Connective/Epithelial 2349 HOUEF84 Cancer 2350 HOUEJ43 Connective/Epithelial 2351 HOUGS36 Connective/Epithelial 2352 HOUHQ36 Connective/Epithelial 2353 HOUIG68 Cancer 2354 HOUIG92 Cancer 2355 HOVAD93 Reproductive 2356 HOVAE10 Cancer 2357 HOVAE36 Reproductive 2358 HOVAE82 Immune/Hematopoietic, Reproductive 2359 HOVAJ68 Reproductive 2360 HOVAW46 Musculoskeletal, Reproductive 2361 HOVBB19 Musculoskeletal, Reproductive 2362 HOVBD31 Cancer 2363 HOVBE81 Reproductive 2364 HOVBI16 Cancer 2365 HOVBS68 Cancer 2366 HOVCC73 Immune/Hematopoietic, Reproductive 2367 HOVCF30 Immune/Hematopoietic, Reproductive 2368 HOVCJ71 Reproductive 2369 HOVCN53 Reproductive 2370 HOVCO53 Reproductive 2371 HPASF94 Cancer 2372 HPBCG26 Cancer 2373 HPBCT11 Reproductive 2374 HPBDE33 Cancer 2375 HPBDE33 Cancer 2376 HPBDF31 Cancer 2377 HPCAG17 Immune/Hematopoietic, Reproductive 2378 HPCAG17 Immune/Hematopoietic, Reproductive 2379 HPCAM02 Immune/Hematopoietic, Musculoskeletal, Reproductive 2380 HPDDQ17 Endocrine 2381 HPDDQ28 Endocrine, Musculoskeletal 2382 HPDDT14 Cancer 2383 HPEAA65 Digestive, Immune/Hematopoietic, Reproductive 2384 HPEAG24 Cancer 2385 HPEBA84 Immune/Hematopoietic, Reproductive 2386 HPEBF91 Cancer 2387 HPEBI09 Digestive, Reproductive 2388 HPFCJ75 Cancer 2389 HPFCP75 Immune/Hematopoietic, Neural/Sensory, Reproductive 2390 HPFDB66 Cancer 2391 HPFDD28 Reproductive 2392 HPFDI47 Digestive, Reproductive 2393 HPIAF35 Reproductive 2394 HPIAK27 Cancer 2395 HPIAL55 Cancer 2396 HPIAT18 Cancer 2397 HPIAZ52 Reproductive 2398 HPIBA07 Cancer 2399 HPIBA24 Cancer 2400 HPIBI40 Cancer 2401 HPIBT19 Reproductive 2402 HPJAA82 Reproductive 2403 HPJAB75 Cancer 2404 HPJAN76 Cancer 2405 HPJAN76 Cancer 2406 HPJAU94 Immune/Hematopoietic, Reproductive 2407 HPJAW78 Immune/Hematopoietic, Musculoskeletal, Reproductive 2408 HPJBS16 Connective/Epithelial, Reproductive 2409 HPJBU04 Immune/Hematopoietic, Reproductive 2410 HPJCN83 Reproductive 2411 HPJCP75 Reproductive 2412 HPJCV35 Reproductive 2413 HPJCX13 Cancer 2414 HPLAW13 Cancer 2415 HPMAI31 Cancer 2416 HPMBI91 Reproductive 2417 HPMBT05 Reproductive 2418 HPMBW95 Cancer 2419 HPMCW10 Cancer 2420 HPMCZ18 Cancer 2421 HPMDA80 Cancer 2422 HPMDD27 Cancer 2423 HPMDF45 Excretory, Immune/Hematopoietic, Reproductive 2424 HPMDP57 Immune/Hematopoietic, Musculoskeletal, Reproductive 2425 HPMEG72 Cancer 2426 HPMFM70 Immune/Hematopoietic, Neural/Sensory, Reproductive 2427 HPMFP48 Cancer 2428 HPMFW01 Cancer 2429 HPMGM06 Digestive, Neural/Sensory, Reproductive 2430 HPMGW43 Cancer 2431 HPMGY89 Cancer 2432 HPMKB09 Cancer 2433 HPMSH26 Cancer 2434 HPMSH96 Mixed Fetal, Reproductive 2435 HPQAJ25 Cardiovascular, Digestive, Mixed Fetal 2436 HPQAJ27 Cancer 2437 HPQAN50 Reproductive 2438 HPQAO80 Cancer 2439 HPQAW27 Cancer 2440 HPQBC90 Cancer 2441 HPQBJ48 Cancer 2442 HPQBJ48 Cancer 2443 HPQBL67 Cancer 2444 HPQBT17 Cancer 2445 HPQCF94 Cancer 2446 HPQCI62 Cancer 2447 HPQRS74 Cancer 2448 HPRAD30 Cancer 2449 HPRCC91 Cancer 2450 HPRCF40 Cancer 2451 HPRCF50 Cancer 2452 HPRCL58 Reproductive 2453 HPRCM72 Cancer 2454 HPRCS59 Reproductive 2455 HPRCT73 Cancer 2456 HPRTH56 Cancer 2457 HPTRE80 Cancer 2458 HPTRI42 Cancer 2459 HPTRL95 Cancer 2460 HPTRQ52 Cancer 2461 HPTTH35 Cancer 2462 HPTTI65 Endocrine, Reproductive 2463 HPTTT62 Cancer 2464 HPTVH24 Cancer 2465 HPTVH59 Endocrine, Neural/Sensory 2466 HPTVI04 Cancer 2467 HPTVI96 Cancer 2468 HPVAA15 Cancer 2469 HPVAB20 Cancer 2470 HPVAB63 Cancer 2471 HPVAF86 Reproductive 2472 HPWAH55 Digestive 2473 HPWAO89 Immune/Hematopoietic, Reproductive 2474 HPWAS27 Cancer 2475 HPWAT86 Immune/Hematopoietic, Neural/Sensory, Reproductive 2476 HPWAV82 Reproductive 2477 HPWBA36 Reproductive 2478 HPWTF23 Cancer 2479 HPWTF23 Cancer 2480 HPWTF53 Cancer 2481 HPXAB56 Immune/Hematopoietic 2482 HPZAB75 Digestive, Reproductive 2483 HRAAB26 Excretory 2484 HRAAC36 Excretory, Immune/Hematopoietic 2485 HRAAF59 Excretory 2486 HRAAG89 Cancer 2487 HRAAO40 Excretory 2488 HRAAZ12 Cancer 2489 HRABA19 Excretory, Reproductive 2490 HRABP28 Excretory, Immune/Hematopoietic 2491 HRABU56 Cardiovascular, Excretory, Musculoskeletal 2492 HRABZ80 Excretory, Immune/Hematopoietic, Musculoskeletal 2493 HRACB01 Excretory 2494 HRACI39 Excretory 2495 HRADU15 Excretory 2496 HRDAH04 Immune/Hematopoietic, Mixed Fetal, Musculoskeletal 2497 HRDBA20 Musculoskeletal 2498 HRDBD32 Musculoskeletal 2499 HRDBL01 Mixed Fetal, Musculoskeletal, Neural/Sensory 2500 HRDDM85 Musculoskeletal 2501 HRDDS22 Cancer 2502 HRDEJ86 Cancer 2503 HRDEQ34 Musculoskeletal 2504 HRDFE30 Cancer 2505 HRDFT83 Musculoskeletal 2506 HRGCA01 Musculoskeletal 2507 HRGCA06 Cancer 2508 HRGSE38 Cancer 2509 HRLAT43 Cancer 2510 HRLME03 Cancer 2511 HROAN20 Cardiovascular, Digestive 2512 HROAP64 Digestive 2513 HROAS35 Digestive 2514 HROAY16 Digestive 2515 HROBJ10 Cancer 2516 HROBW46 Digestive, Immune/Hematopoietic 2517 HRODG86 Cancer 2518 HRSAL26 Cancer 2519 HRTAE88 Digestive 2520 HRTAP63 Cancer 2521 HRTAR24 Digestive, Immune/Hematopoietic 2522 HSAAN03 Cancer 2523 HSAAS05 Immune/Hematopoietic, Neural/Sensory 2524 HSAAW13 Cancer 2525 HSABA15 Cancer 2526 HSABG81 Cancer 2527 HSATA50 Cardiovascular, Immune/Hematopoietic, Musculoskeletal 2528 HSATA61 Cancer 2529 HSATG66 Cancer 2530 HSATI91 Immune/Hematopoietic 2531 HSATR50 Digestive, Immune/Hematopoietic, Reproductive 2532 HSATT82 Immune/Hematopoietic 2533 HSATW19 Immune/Hematopoietic, Musculoskeletal 2534 HSATW67 Excretory, Immune/Hematopoietic, Reproductive 2535 HSATZ02 Immune/Hematopoietic, Musculoskeletal 2536 HSAUA95 Immune/Hematopoietic 2537 HSAUB89 Cancer 2538 HSAUI53 Immune/Hematopoietic 2539 HSAUV74 Cancer 2540 HSAUX39 Immune/Hematopoietic 2541 HSAVA58 Immune/Hematopoietic, Mixed Fetal 2542 HSAVE52 Immune/Hematopoietic 2543 HSAVH32 Immune/Hematopoietic 2544 HSAVM49 Immune/Hematopoietic 2545 HSAVO11 Immune/Hematopoietic, Neural/Sensory 2546 HSAVO17 Immune/Hematopoietic, Musculoskeletal 2547 HSAVQ13 Immune/Hematopoietic 2548 HSAVR85 Cancer 2549 HSAVY92 Immune/Hematopoietic, Neural/Sensory 2550 HSAVZ05 Digestive, Immune/Hematopoietic 2551 HSAWB58 Immune/Hematopoietic 2552 HSAWH36 Immune/Hematopoietic 2553 HSAWM20 Immune/Hematopoietic 2554 HSAWM74 Cancer 2555 HSAWX70 Cancer 2556 HSAXC22 Immune/Hematopoietic 2557 HSAXI10 Digestive, Immune/Hematopoietic 2558 HSAXL49 Immune/Hematopoietic 2559 HSAXL82 Immune/Hematopoietic 2560 HSAXN57 Immune/Hematopoietic 2561 HSAXO45 Immune/Hematopoietic 2562 HSAXS06 Immune/Hematopoietic, Reproductive 2563 HSAXS22 Immune/Hematopoietic 2564 HSAYL24 Immune/Hematopoietic 2565 HSAYO82 Endocrine, Immune/Hematopoietic 2566 HSAYR62 Cancer 2567 HSAZP90 Immune/Hematopoietic 2568 HSBAJ47 Cancer 2569 HSDBI90 Digestive, Endocrine, Neural/Sensory 2570 HSDDC55 Neural/Sensory 2571 HSDEA26 Neural/Sensory 2572 HSDEY39 Neural/Sensory 2573 HSDFF72 Cancer 2574 HSDFO08 Neural/Sensory 2575 HSDFR10 Digestive, Neural/Sensory, Reproductive 2576 HSDGB20 Neural/Sensory 2577 HSDGH56 Cancer 2578 HSDGM01 Neural/Sensory 2579 HSDGM42 Cancer 2580 HSDGM42 Cancer 2581 HSDGM42 Cancer 2582 HSDGM42 Cancer 2583 HSDGM42 Cancer 2584 HSDGM42 Cancer 2585 HSDHD05 Neural/Sensory 2586 HSDIE51 Cancer 2587 HSDIK31 Cancer 2588 HSDIV37 Cancer 2589 HSDJC96 Cancer 2590 HSDJE77 Cardiovascular, Immune/Hematopoietic, Neural/Sensory 2591 HSDJF04 Cancer 2592 HSDJG47 Cancer 2593 HSDJH72 Connective/Epithelial, Excretory, Neural/Sensory 2594 HSDJL07 Neural/Sensory, Reproductive 2595 HSDJR49 Neural/Sensory 2596 HSDJV24 Cancer 2597 HSDJV40 Immune/Hematopoietic, Neural/Sensory 2598 HSDKA64 Immune/Hematopoietic, Neural/Sensory 2599 HSDKE82 Neural/Sensory 2600 HSDKF96 Neural/Sensory 2601 HSDZO08 Cancer 2602 HSDZQ96 Neural/Sensory 2603 HSEBB18 Cancer 2604 HSFAM19 Cancer 2605 HSHAG54 Cancer 2606 HSHAS72 Cancer 2607 HSHAX04 Cancer 2608 HSHBT15 Cancer 2609 HSHCE85 Cancer 2610 HSIAC81 Digestive 2611 HSIAF66 Digestive 2612 HSIAP01 Digestive, Reproductive 2613 HSIDA33 Cancer 2614 HSIDA39 Digestive 2615 HSIDZ25 Cancer 2616 HSIEB64 Digestive 2617 HSIEM18 Cancer 2618 HSIFO61 Cancer 2619 HSIFO61 Cancer 2620 HSIGC63 Digestive, Immune/Hematopoietic, Reproductive 2621 HSIGM95 Digestive, Immune/Hematopoietic 2622 HSJAE76 Cancer 2623 HSJAN83 Digestive, Musculoskeletal 2624 HSJAQ10 Cancer 2625 HSJAR59 Musculoskeletal 2626 HSJAU93 Cancer 2627 HSJAY14 Cancer 2628 HSJAY14 Cancer 2629 HSJAY14 Cancer 2630 HSJAY14 Cancer 2631 HSJBB27 Musculoskeletal 2632 HSKBU03 Musculoskeletal, Neural/Sensory 2633 HSKCQ51 Cancer 2634 HSKDE13 Cancer 2635 HSKDS47 Cancer 2636 HSKHV81 Musculoskeletal 2637 HSKXB14 Cancer 2638 HSKYR49 Cancer 2639 HSKYU81 Cancer 2640 HSKYY92 Musculoskeletal 2641 HSLAB11 Cancer 2642 HSLAS96 Immune/Hematopoietic, Musculoskeletal 2643 HSLAW59 Immune/Hematopoietic, Musculoskeletal 2644 HSLCH54 Cancer 2645 HSLCH57 Cancer 2646 HSLCI86 Endocrine, Mixed Fetal, Musculoskeletal 2647 HSLCS31 Cancer 2648 HSLCS34 Cancer 2649 HSLCV16 Cancer 2650 HSLDW54 Cancer 2651 HSLEC18 Cancer 2652 HSLEG59 Musculoskeletal 2653 HSLFR59 Cancer 2654 HSLGD91 Cancer 2655 HSLGF66 Cancer 2656 HSLGF70 Musculoskeletal, Neural/Sensory 2657 HSLGP68 Musculoskeletal, Neural/Sensory 2658 HSNAB88 Cancer 2659 HSNAH56 Cancer 2660 HSNAN38 Cancer 2661 HSNAO19 Cancer 2662 HSNAQ52 Cancer 2663 HSNAT08 Cancer 2664 HSNAW06 Immune/Hematopoietic 2665 HSNAW37 Cancer 2666 HSNBJ05 Cancer 2667 HSNBO90 Cancer 2668 HSNBQ36 Cancer 2669 HSNBS39 Cancer 2670 HSOAE34 Digestive, Immune/Hematopoietic 2671 HSOAT44 Cancer 2672 HSOBB94 Cancer 2673 HSOBH11 Digestive 2674 HSOBP75 Cancer 2675 HSOBW65 Digestive 2676 HSPAA89 Digestive 2677 HSPAC13 Cancer 2678 HSPAG75 Digestive 2679 HSPAI20 Digestive, Neural/Sensory 2680 HSPAL59 Digestive, Immune/Hematopoietic 2681 HSPAY90 Cancer 2682 HSPMF63 Cancer 2683 HSQAC69 Cancer 2684 HSQAH14 Cancer 2685 HSQAX94 Cancer 2686 HSQBL20 Cancer 2687 HSQCQ45 Cancer 2688 HSQCY74 Cancer 2689 HSQDM74 Cancer 2690 HSQEG23 Cancer 2691 HSQEG47 Cancer 2692 HSQFE72 Cancer 2693 HSQFE76 Cancer 2694 HSQFV12 Cancer 2695 HSRAA81 Cancer 2696 HSRAO56 Cancer 2697 HSRAV28 Digestive, Musculoskeletal 2698 HSRDM56 Cancer 2699 HSRDW57 Cancer 2700 HSREC72 Immune/Hematopoietic, Musculoskeletal 2701 HSREG42 Cancer 2702 HSRFD18 Cancer 2703 HSRGZ11 Cancer 2704 HSRHB59 Cancer 2705 HSSAN03 Cancer 2706 HSSCC66 Musculoskeletal 2707 HSSDI13 Musculoskeletal 2708 HSSDQ20 Musculoskeletal, Neural/Sensory 2709 HSSDX38 Musculoskeletal 2710 HSSED57 Cancer 2711 HSSEL28 Cancer 2712 HSSFP88 Cancer 2713 HSSGS62 Musculoskeletal, Reproductive 2714 HSSJA23 Cancer 2715 HSSJF26 Musculoskeletal 2716 HSSJF96 Musculoskeletal 2717 HSSJM47 Cancer 2718 HSSJW30 Cancer 2719 HSSJW30 Cancer 2720 HSSJW30 Cancer 2721 HSSMY35 Cancer 2722 HSTAL93 Connective/Epithelial 2723 HSTBG23 Connective/Epithelial 2724 HSUAF06 Immune/Hematopoietic 2725 HSUBX67 Immune/Hematopoietic 2726 HSUSB73 Immune/Hematopoietic, Reproductive 2727 HSVAC05 Cancer 2728 HSVAE42 Connective/Epithelial, Neural/Sensory 2729 HSVAL83 Cancer 2730 HSVAT36 Immune/Hematopoietic 2731 HSVAV02 Cancer 2732 HSVBA83 Endocrine, Mixed Fetal 2733 HSVBD37 Cancer 2734 HSVBN46 Cancer 2735 HSVBY62 Cancer 2736 HSVBZ53 Cancer 2737 HSVCF53 Cardiovascular, Neural/Sensory, Reproductive 2738 HSWAZ17 Connective/Epithelial, Reproductive, Respiratory 2739 HSWBI16 Cancer 2740 HSXAI44 Neural/Sensory 2741 HSXAJ07 Neural/Sensory 2742 HSXAS59 Neural/Sensory 2743 HSXAX20 Digestive, Mixed Fetal, Neural/Sensory 2744 HSXAY60 Cancer 2745 HSXBB78 Neural/Sensory 2746 HSXCA83 Cancer 2747 HSXCX20 Cancer 2748 HSXFG21 Cancer 2749 HSXFH82 Cancer 2750 HSYBD33 Immune/Hematopoietic 2751 HSYBR79 Cancer 2752 HSYBV44 Immune/Hematopoietic 2753 HSYBZ94 Cancer 2754 HT3AB13 Cancer 2755 HT4SB02 Immune/Hematopoietic 2756 HT4SB37 Cardiovascular, Immune/Hematopoietic, Reproductive 2757 HT4SB81 Cancer 2758 HT4SB81 Cancer 2759 HT4SB81 Cancer 2760 HT5FX76 Cancer 2761 HTABF81 Cancer 2762 HTACX63 Immune/Hematopoietic 2763 HTADC63 Cancer 2764 HTADO61 Cancer 2765 HTADQ22 Cancer 2766 HTAEC59 Cancer 2767 HTAED89 Immune/Hematopoietic 2768 HTAEF02 Immune/Hematopoietic 2769 HTAEH58 Immune/Hematopoietic 2770 HTAEO35 Immune/Hematopoietic 2771 HTDAF68 Immune/Hematopoietic 2772 HTDAI38 Cancer 2773 HTEAJ87 Mixed Fetal, Neural/Sensory, Reproductive 2774 HTEAN76 Cancer 2775 HTEBL56 Cancer 2776 HTECE87 Cancer 2777 HTEDF78 Reproductive 2778 HTEDT87 Cancer 2779 HTEDX05 Cancer 2780 HTEEC19 Cancer 2781 HTEGH03 Cancer 2782 HTEGH03 Cancer 2783 HTEGS48 Reproductive 2784 HTEGY81 Cancer 2785 HTEHB11 Reproductive 2786 HTEHB49 Immune/Hematopoietic, Reproductive 2787 HTEHS91 Cancer 2788 HTEHV60 Reproductive 2789 HTEHW80 Reproductive 2790 HTEID25 Reproductive 2791 HTEIJ23 Cancer 2792 HTEIM62 Digestive, Immune/Hematopoietic, Reproductive 2793 HTEIV33 Reproductive 2794 HTEIV65 Reproductive 2795 HTEJC50 Reproductive 2796 HTEJD20 Cancer 2797 HTEJD61 Reproductive 2798 HTEJF31 Reproductive 2799 HTEJI29 Reproductive 2800 HTEJL16 Reproductive 2801 HTEJP65 Cancer 2802 HTEJY20 Cancer 2803 HTEKD35 Reproductive 2804 HTEKP82 Cardiovascular, Mixed Fetal, Reproductive 2805 HTEKV69 Reproductive 2806 HTEKZ52 Reproductive 2807 HTEQG28 Immune/Hematopoietic, Reproductive 2808 HTFOB75 Cancer 2809 HTGAA35 Immune/Hematopoietic 2810 HTGAD74 Immune/Hematopoietic, Reproductive 2811 HTGAP05 Immune/Hematopoietic, Neural/Sensory 2812 HTGAQ29 Immune/Hematopoietic 2813 HTGAR21 Immune/Hematopoietic 2814 HTGAS70 Cancer 2815 HTGAT65 Immune/Hematopoietic, Neural/Sensory, Reproductive 2816 HTGAU17 Immune/Hematopoietic 2817 HTGBF47 Immune/Hematopoietic 2818 HTGBK95 Cancer 2819 HTGCC01 Immune/Hematopoietic 2820 HTGCK43 Cancer 2821 HTGDS43 Immune/Hematopoietic, Neural/Sensory 2822 HTGDS92 Cancer 2823 HTGEX34 Digestive, Immune/Hematopoietic 2824 HTGFM31 Immune/Hematopoietic 2825 HTGGM37 Digestive, Immune/Hematopoietic 2826 HTGGN22 Immune/Hematopoietic 2827 HTHAA41 Cancer 2828 HTHBC58 Digestive, Immune/Hematopoietic 2829 HTHBO72 Cancer 2830 HTHBQ29 Immune/Hematopoietic 2831 HTHBT76 Cancer 2832 HTHBZ91 Immune/Hematopoietic 2833 HTHCA30 Cancer 2834 HTHCM60 Immune/Hematopoietic 2835 HTHDB20 Immune/Hematopoietic 2836 HTHDF45 Immune/Hematopoietic 2837 HTHDF86 Immune/Hematopoietic 2838 HTHDH18 Immune/Hematopoietic 2839 HTHDP65 Cancer 2840 HTHDT25 Immune/Hematopoietic 2841 HTHDV50 Immune/Hematopoietic 2842 HTJMA64 Cancer 2843 HTJMJ72 Connective/Epithelial, Immune/Hematopoietic 2844 HTLAD74 Reproductive 2845 HTLAF81 Cancer 2846 HTLBF46 Cancer 2847 HTLBF63 Cancer 2848 HTLCX82 Cancer 2849 HTLDD89 Reproductive 2850 HTLDN34 Reproductive 2851 HTLDP19 Cancer 2852 HTLDY30 Cancer 2853 HTLEJ24 Cancer 2854 HTLEJ75 Cancer 2855 HTLEJ75 Cancer 2856 HTLEP55 Cancer 2857 HTLEV80 Cancer 2858 HTLEZ57 Cancer 2859 HTLFA90 Cancer 2860 HTLGL33 Reproductive 2861 HTLGQ25 Reproductive 2862 HTLGS72 Reproductive 2863 HTLGY50 Cancer 2864 HTLHN86 Cancer 2865 HTLHN86 Cancer 2866 HTLHN86 Cancer 2867 HTLHN86 Cancer 2868 HTLIW29 Cancer 2869 HTLJC15 Cancer 2870 HTNAL14 Cancer 2871 HTNAL34 Endocrine, Reproductive 2872 HTNBJ15 Cancer 2873 HTNBJ15 Cancer 2874 HTNBJ15 Cancer 2875 HTNBJ15 Cancer 2876 HTOAC65 Immune/Hematopoietic 2877 HTOAE47 Immune/Hematopoietic 2878 HTOAK03 Cancer 2879 HTOAO58 Immune/Hematopoietic 2880 HTOAT56 Cancer 2881 HTOBG07 Immune/Hematopoietic, Musculoskeletal 2882 HTOBG62 Immune/Hematopoietic 2883 HTODA92 Cancer 2884 HTODN35 Immune/Hematopoietic 2885 HTODO45 Immune/Hematopoietic 2886 HTOEA53 Digestive, Excretory, Immune/Hematopoietic 2887 HTOEB55 Cancer 2888 HTOEB76 Immune/Hematopoietic 2889 HTOET03 Cancer 2890 HTOET03 Cancer 2891 HTOEV01 Immune/Hematopoietic, Reproductive 2892 HTOFA11 Cancer 2893 HTOFC33 Immune/Hematopoietic 2894 HTOGB79 Cancer 2895 HTOHE22 Immune/Hematopoietic 2896 HTOHG63 Cancer 2897 HTOHJ93 Immune/Hematopoietic 2898 HTOHM12 Immune/Hematopoietic, Neural/Sensory 2899 HTOHM82 Cancer 2900 HTOHN40 Immune/Hematopoietic, Neural/Sensory 2901 HTOHR59 Digestive, Immune/Hematopoietic, Neural/Sensory 2902 HTOHS29 Cancer 2903 HTOID65 Cancer 2904 HTOIE17 Excretory, Immune/Hematopoietic 2905 HTOIG16 Immune/Hematopoietic, Reproductive 2906 HTOIH39 Immune/Hematopoietic 2907 HTOIH51 Immune/Hematopoietic 2908 HTOJB02 Immune/Hematopoietic 2909 HTOJJ26 Connective/Epithelial, Digestive, Immune/Hematopoietic 2910 HTOJP25 Immune/Hematopoietic 2911 HTOJS23 Immune/Hematopoietic 2912 HTOJY56 Cancer 2913 HTOJZ18 Immune/Hematopoietic 2914 HTPCG10 Cancer 2915 HTPCO75 Cancer 2916 HTPCW21 Digestive, Neural/Sensory 2917 HTPDD68 Cancer 2918 HTPDV75 Digestive, Reproductive 2919 HTSER28 Cancer 2920 HTSET62 Cancer 2921 HTSFV18 Cancer 2922 HTSGO13 Cancer 2923 HTSGO88 Immune/Hematopoietic 2924 HTTAH05 Reproductive 2925 HTTAP37 Immune/Hematopoietic, Reproductive 2926 HTTBJ38 Cancer 2927 HTTDB11 Cancer 2928 HTTDG27 Reproductive 2929 HTTDN24 Cancer 2930 HTTDO33 Cancer 2931 HTTDT67 Cancer 2932 HTTEO25 Cancer 2933 HTTEP11 Neural/Sensory, Reproductive 2934 HTTES77 Cancer 2935 HTTFD29 Reproductive 2936 HTTFG15 Cancer 2937 HTWAM19 Immune/Hematopoietic 2938 HTWBF58 Immune/Hematopoietic 2939 HTWBO30 Cancer 2940 HTWBZ57 Cancer 2941 HTWCC10 Immune/Hematopoietic 2942 HTWCE14 Cancer 2943 HTWCT76 Digestive, Immune/Hematopoietic 2944 HTWDJ17 Cancer 2945 HTWDM89 Immune/Hematopoietic 2946 HTWEA05 Immune/Hematopoietic 2947 HTWEG06 Immune/Hematopoietic 2948 HTWEQ36 Cancer 2949 HTWFA21 Immune/Hematopoietic 2950 HTWFA88 Digestive, Immune/Hematopoietic 2951 HTWFM85 Cancer 2952 HTWFO43 Cancer 2953 HTWLG39 Cancer 2954 HTXAA20 Cancer 2955 HTXAD75 Cancer 2956 HTXAR92 Immune/Hematopoietic 2957 HTXBS38 Cancer 2958 HTXBU88 Immune/Hematopoietic 2959 HTXCP27 Cancer 2960 HTXCU30 Excretory, Immune/Hematopoietic 2961 HTXCV44 Immune/Hematopoietic, Neural/Sensory 2962 HTXDJ21 Immune/Hematopoietic 2963 HTXDJ75 Digestive, Immune/Hematopoietic, Mixed Fetal 2964 HTXDJ85 Immune/Hematopoietic 2965 HTXDK09 Cancer 2966 HTXDO17 Immune/Hematopoietic, Neural/Sensory, Respiratory 2967 HTXDT72 Cancer 2968 HTXDU08 Cancer 2969 HTXDZ68 Immune/Hematopoietic, Musculoskeletal 2970 HTXEN33 Immune/Hematopoietic, Reproductive 2971 HTXES13 Cancer 2972 HTXFD86 Cancer 2973 HTXGK12 Cancer 2974 HTXGL32 Immune/Hematopoietic 2975 HTXJD08 Digestive, Immune/Hematopoietic 2976 HTXJD85 Immune/Hematopoietic 2977 HTXJE12 Cancer 2978 HTXJI59 Cancer 2979 HTXJJ92 Cancer 2980 HTXJM94 Cancer 2981 HTXJV54 Digestive, Immune/Hematopoietic, Reproductive 2982 HTXJW06 Cancer 2983 HTXKB57 Cancer 2984 HTXKH22 Immune/Hematopoietic 2985 HTXKH40 Cancer 2986 HTXKK76 Immune/Hematopoietic 2987 HTXKL53 Cancer 2988 HTXKS11 Immune/Hematopoietic 2989 HTXKS27 Cancer 2990 HTXLC05 Digestive, Immune/Hematopoietic, Respiratory 2991 HTXLC45 Immune/Hematopoietic 2992 HTXLT36 Cancer 2993 HTXLY94 Cancer 2994 HTXNV66 Cancer 2995 HTXOL30 Immune/Hematopoietic 2996 HTXOW27 Cancer 2997 HTXPD86 Cancer 2998 HTXPT57 Digestive, Immune/Hematopoietic 2999 HTYSJ88 Endocrine, Immune/Hematopoietic 3000 HUDBE20 Reproductive 3001 HUDBK47 Immune/Hematopoietic, Reproductive 3002 HUFAB57 Cancer 3003 HUFAL17 Digestive 3004 HUFAO92 Digestive, Reproductive 3005 HUFAO94 Cancer 3006 HUFAP33 Cancer 3007 HUFAU71 Cancer 3008 HUFBK95 Digestive, Reproductive 3009 HUFBP77 Cancer 3010 HUFBV62 Cancer 3011 HUFBY96 Cancer 3012 HUFCN72 Digestive 3013 HUFEF79 Cancer 3014 HUKAD46 Endocrine, Immune/Hematopoietic, Reproductive 3015 HUKAI28 Cardiovascular, Reproductive 3016 HUKAO50 Cancer 3017 HUKCS86 Cancer 3018 HUKCS86 Cancer 3019 HUKEA22 Cancer 3020 HUKEL79 Cancer 3021 HUKEX37 Reproductive 3022 HUKFC71 Cancer 3023 HUKFV37 Cancer 3024 HUKFY09 Cancer 3025 HUNAL39 Reproductive 3026 HUSAO04 Cancer 3027 HUSAO04 Cancer 3028 HUSCA09 Cancer 3029 HUSCJ01 Cancer 3030 HUSGB23 Cancer 3031 HUSGJ09 Cardiovascular, Neural/Sensory 3032 HUSGQ57 Cancer 3033 HUSGY15 Cancer 3034 HUSHD41 Cancer 3035 HUSHK65 Cancer 3036 HUSIK45 Cancer 3037 HUSIO57 Cancer 3038 HUSIP17 Cardiovascular 3039 HUSIR70 Cancer 3040 HUSXP50 Cardiovascular, Reproductive 3041 HUSXY93 Cancer 3042 HUSYG26 Cancer 3043 HUVCQ68 Cancer 3044 HUVDG58 Digestive, Immune/Hematopoietic, Reproductive 3045 HUVEG53 Cancer 3046 HWAAH11 Cancer 3047 HWAAQ28 Cancer 3048 HWAAY60 Cancer 3049 HWABR43 Digestive, Immune/Hematopoietic 3050 HWACH06 Cancer 3051 HWACZ33 Digestive, Immune/Hematopoietic, Reproductive 3052 HWADV90 Immune/Hematopoietic 3053 HWAEB52 Cancer 3054 HWBAK71 Immune/Hematopoietic 3055 HWBBU75 Cancer 3056 HWBCN81 Immune/Hematopoietic, Reproductive 3057 HWBCP16 Immune/Hematopoietic 3058 HWBCX93 Cancer 3059 HWBEF34 Immune/Hematopoietic, Neural/Sensory 3060 HWFBB23 Cancer 3061 HWFBI40 Connective/Epithelial, Digestive, Immune/Hematopoietic 3062 HWHGV77 Connective/Epithelial 3063 HWHGW09 Cancer 3064 HWHHA21 Connective/Epithelial 3065 HWHPU44 Connective/Epithelial 3066 HWHRC51 Cancer 3067 HWLAT50 Cancer 3068 HWLBO67 Digestive 3069 HWLGP26 Cancer 3070 HWLHO31 Cardiovascular, Digestive 3071 HWLIL31 Cancer 3072 HWLJN08 Cancer 3073 HWLRE03 Cancer 3074 HWTAM38 Digestive, Immune/Hematopoietic, Reproductive 3075 HWTAW58 Cancer 3076 HWTBB42 Cancer 3077 HWTBC75 Cancer 3078 HWTBI25 Cancer 3079 HWTBL86 Cancer 3080 HWTBX66 Cancer 3081 HYAAC74 Immune/Hematopoietic, Musculoskeletal, Reproductive 3082 HYAAD61 Immune/Hematopoietic 3083 HYACC21 Immune/Hematopoietic 3084 HYBAP75 Cancer 3085 HYBAQ24 Cancer 3086 HYBAW56 Musculoskeletal 3087 HYBBD81 Musculoskeletal

Table 1E provides information related to biological activities and preferred indications for polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof). Table 1E also provides information related to assays which may be used to test polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) for the corresponding biological activities. The first column (“Gene No.”) provides the gene number in the application for each clone identifier. The second column (“cDNA Clone ID:”) provides the unique clone identifier for each clone as previously described and indicated in Tables 1A, 1B, 1C, and 1D. The third column (“AA SEQ ID NO:Y”) indicates the Sequence Listing SEQ ID Number for polypeptide sequences encoded by the corresponding cDNA clones (also as indicated in Tables 1A, 1B, and 2). The fourth column (“Biological Activity”) indicates a biological activity corresponding to the indicated polypeptides (or polynucleotides encoding said polypeptides). The fifth column (“Exemplary Activity Assay”) further describes the corresponding biological activity and also provides information pertaining to the various types of assays which may be performed to test, demonstrate, or quantify the corresponding biological activity. The sixth column (“Preferred Indictions”) describes particular embodiments of the invention as well as indications (e.g. pathologies, diseases, disorders, abnormalities, etc.) for which polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) may be used in detecting, diagnosing, preventing, and/or treating.

Table 1E describes the use of, inter alia, FMAT technology for testing or demonstrating various biological activities. Fluorometric microvolume assay technology (FMAT) is a fluorescence-based system which provides a means to perform nonradioactive cell- and bead-based assays to detect activation of cell signal transduction pathways. This technology was designed specifically for ligand binding and immunological assays. Using this technology, fluorescent cells or beads at the bottom of the well are detected as localized areas of concentrated fluorescence using a data processing system. Unbound flurophore comprising the background signal is ignored, allowing for a wide variety of homogeneous assays. FMAT technology may be used for peptide ligand binding assays, immunofluorescence, apoptosis, cytotoxicity, and bead-based immunocapture assays. See, Miraglia S et. al., “Homogeneous cell and bead based assays for highthroughput screening using flourometric microvolume assay technology,” Journal of Biomolecular Screening; 4:193-204 (1999). In particular, FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides (including polypeptide fragments and variants) to activate signal transduction pathways. For example, FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides to upregulate production of immunomodulatory proteins (such as, for example, interleukins, GM-CSF, Rantes, and Tumor Necrosis factors, as well as other cellular regulators (e.g. insulin)).

Table 1E also describes the use of kinase assays for testing, demonstrating, or quantifying biological activity. In this regard, the phosphorylation and de-phosphorylation of specific amino acid residues (e.g. Tyrosine, Serine, Threonine) on cell-signal transduction proteins provides a fast, reversible means for activation and de-activation of cellular signal transduction pathways. Moreover, cell signal transduction via phosphorylation/de-phosphorylation is crucial to the regulation of a wide variety of cellular processes (e.g. proliferation, differentiation, migration, apoptosis, etc.). Accordingly, kinase assays provide a powerful tool useful for testing, confirming, and/or identifying polypeptides (including polypeptide fragments and variants) that mediate cell signal transduction events via protein phosphorylation. See e.g., Forrer, P., Tamaskovic R., and Jaussi, R. “Enzyme-Linked Immunosorbent Assay for Measurement of JNK, ERK, and p38 Kinase Activities” Biol. Chem. 379(8-9): 1101-1110 (1998). LENGTHY TABLE REFERENCED HERE US20070032413A1-20070208-T00003 Please refer to the end of the specification for access instructions. Table 1F:

Polynucleotides encoding polypeptides of the present invention can be used in assays to test for one or more biological activities. One such biological activity which may be tested includes the ability of polynucleotides and polypeptides of the invention to stimulate up-regulation or down-regulation of expression of particular genes and proteins. Hence, if polynucleotides and polypeptides of the present invention exhibit activity in altering particular gene and protein expression patterns, it is likely that these polynucleotides and polypeptides of the present invention may be involved in, or capable of effecting changes in, diseases associated with the altered gene and protein expression profiles. Hence, polynucleotides, polypeptides, or antibodies of the present invention could be used to treat said associated diseases.

TaqMan® assays may be performed to assess the ability of polynucleotides (and polypeptides they encode) to alter the expression pattern of particular “target” genes. TaqMan® reactions are performed to evaluate the ability of a test agent to induce or repress expression of specific genes in different cell types. TaqMan® gene expression quantification assays (“TaqMan® assays”) are well known to, and routinely performed by, those of ordinary skill in the art. TaqMan® assays are performed in a two step reverse transcription/polymerase chain reaction (RT-PCR). In the first (RT) step, cDNA is reverse transcribed from total RNA samples using random hexamer primers. In the second (PCR) step, PCR products are synthesized from the cDNA using gene specific primers.

To quantify gene expression the Taqman® PCR reaction exploits the 5′ nuclease activity of AmpliTaq Gold® DNA Polymerase to cleave a Taqman® probe (distinct from the primers) during PCR. The Taqman® probe contains a reporter dye at the 5′-end of the probe and a quencher dye at the 3′ end of the probe. When the probe is intact, the proximity of the reporter dye to the quencher dye results in suppression of the reporter fluorescence. During PCR, if the target of interest is present, the probe specifically anneals between the forward and reverse primer sites. AmpliTaq Fold DNA Polymerase then cleaves the probe between the reporter and quencher when the probe hybridizes to the target, resulting in increased fluorescence of the reporter (see FIG. 2). Accumulation of PCR products is detected directly by monitoring the increase in fluorescence of the reporter dye.

After the probe fragments are displaced from the target, polymerization of the strand continues. The 3′-end of the probe is blocked to prevent extension of the probe during PCR. This process occurs in every cycle and does not interfere with the exponential accumulation of product. The increase in fluorescence signal is detected only if the target sequence is complementary to the probe and is amplified during PCR. Because of these requirements, any nonspecific amplification is not detected.

For test sample preparation, vector controls or constructs containing the coding sequence for the gene of interest are transfected into cells, such as for example 293T cells, and supernatants collected after 48 hours. For cell treatment and RNA isolation, multiple primary human cells or human cell lines are used; such cells may include but are not limited to, Normal Human Dermal Fibroblasts, Aortic Smooth Muscle, Human Umbilical Vein Endothelial Cells, HepG2, Daudi, Jurkat, U937, Caco, and THP-1 cell lines. Cells are plated in growth media and growth is arrested by culturing without media change for 3 days, or by switching cells to low serum media and incubating overnight. Cells are treated for 1, 6, or 24 hours with either vector control supernatant or sample supernatant (or purified/partially purified protein preparations in buffer). Total RNA is isolated; for example, by using Trizol extraction or by using the Ambion RNAqueous™-4PCR RNA isolation system. Expression levels of multiple genes are analyzed using Taqman®, and expression in the test sample is compared to control vector samples to identify genes induced or repressed. Each of the above described techniques are well known to, and routinely performed by, those of ordinary skill in the art.

Table 1F indicates particular disease classes and preferred indications for which polynucleotides, polypeptides, or antibodies of the present invention may be used in detecting, diagnosing, preventing, treating and/or ameliorating said diseases and disorders based on “target” gene expression patterns which may be up- or down-regulated by polynucleotides (and the encoded polypeptides) corresponding to each indicated cDNA Clone ID (shown in Table 1F, Column 2).

Thus, in preferred embodiments, the present invention encompasses a method of detecting, diagnosing, preventing, treating, and/or ameliorating a disease or disorder listed in the “Disease Class” and/or “Preferred Indication” columns of Table 1F; comprising administering to a patient in which such detection, diagnosis, prevention, or treatment is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to detect, diagnose, prevent, treat, or ameliorate the disease or disorder. The first and second columns of Table 1D show the “Gene No.” and “cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may be used in detecting, diagnosing, preventing, treating, or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in the “Disease Class” or “Preferred Indication” Columns of Table 1F.

In another embodiment, the present invention also encompasses methods of detecting, diagnosing, preventing, treating, or ameliorating a disease or disorder listed in the “Disease Class” or “Preferred Indication” Columns of Table 1F; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in the “Disease Class” or “Preferred Indication” Columns of Table 1F.

The “Disease Class” Column of Table 1F provides a categorized descriptive heading for diseases, disorders, and/or conditions (more fully described below) that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The “Preferred Indication” Column of Table 1F describes diseases, disorders, and/or conditions that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The “Cell Line” and “Exemplary Targets” Columns of Table 1F indicate particular cell lines and target genes, respectively, which may show altered gene expression patterns (i.e., up- or down-regulation of the indicated target gene) in Taqman® assays, performed as described above, utilizing polynucleotides of the cDNA Clone ID shown in the corresponding row. Alteration of expression patterns of the indicated “Exemplary Target” genes is correlated with a particular “Disease Class” and/or “Preferred Indication” as shown in the corresponding row under the respective column headings.

The “Exemplary Accessions” Column indicates GenBank Accessions (available online through the National Center for Biotechnology Information (NCBI) at http://www.ncbi.nlm.nih.gov/) which correspond to the “Exemplary Targets” shown in the adjacent row.

The recitation of “Cancer” in the “Disease Class” Column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate neoplastic diseases and/or disorders (e.g., leukemias, cancers, etc., as described below under “Hyperproliferative Disorders”).

The recitation of “Immune” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), blood disorders (e.g., as described below under “Immune Activity” “Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under “Infectious Disease”).

The recitation of “Angiogenesis” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), diseases and/or disorders of the cardiovascular system (e.g., as described below under “Cardiovascular Disorders”), diseases and/or disorders involving cellular and genetic abnormalities (e.g., as described below under “Diseases at the Cellular Level”), diseases and/or disorders involving angiogenesis (e.g., as described below under “Anti-Angiogenesis Activity”), to promote or inhibit cell or tissue regeneration (e.g., as described below under “Regeneration”), or to promote wound healing (e.g., as described below under “Wound Healing and Epithelial Cell Proliferation”).

The recitation of “Diabetes” in the “Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diabetes (including diabetes mellitus types I and II), as well as diseases and/or disorders associated with, or consequential to, diabetes (e.g. as described below under “Endocrine Disorders,” “Renal Disorders,” and “Gastrointestinal Disorders”). TABLE 1F Gene cDNA Disease Exemplary No. Clone ID Class Preferred Indications Cell Line Targets Exemplary Accessions 423 HTEEW69 Immune Highly preferred indications include AOSMC CCR7 gb|X84702|HSDNABLR2 immunological disorders such as described herein CXCR3 gb|Z79783|HSCKRL2 under the heading “Immune Activity” and/or Rag2 gb|AY011962|AY011962 “Blood-Related Disorders” (particularly VLA4 gb|X16983|HSINTAL4 including, but not limited to, immune disorders involving muscle tissues and the cardiovascular system (e.g. heart, lungs, circulatory system)). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving muscle tissue or the cardiovascular system). (AOSMC cells are human aortic smooth muscle cells). 423 HTEEW69 Immune Highly preferred indications include Caco-2 TNF gb|AJ270944|HSA27094 immunological disorders such as described herein under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving the cells of the gastrointestinal tract). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving cells of the gastrointestinal tract). (The Caco-2 cell line is a human colorectal adenocarcinoma cell line available through the ATCC as cell line number HTB-37). 423 HTEEW69 Immune Highly preferred indications include Daudi GATA3 gb|X55037|HSGATA3 immunological disorders such as described herein ICAM gb|X06990|HSICAM1 under the heading “Immune Activity” and/or TNF gb|AJ270944|HSA27094 “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving the B-cells). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving B-cells). (The Daudi cell line is a human B lymphoblast cell line available through the ATCC as cell line number CCL-213). 423 HTEEW69 Immune Highly preferred indications include HEK293 TNF gb|AJ270944|HSA27094 immunological disorders such as described herein under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving epithelial cells or the renal system). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving epithelial cells or the renal system). (The 293 cell line is a human embryonal kidney epithelial cell line available through the ATCC as cell line number CRL-1573). 423 HTEEW69 Immune Highly preferred indications include Liver ICAM gb|X06990|HSICAM1 immunological disorders such as described herein under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving cells of the hepatic system). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving cells of the hepatic system). 423 HTEEW69 Immune Highly preferred indications include NHDF CIS3 gb|AB006967|AB006967 immunological disorders such as described herein TNF gb|AJ270944|HSA27094 under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving the skin). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving the skin). (NHDF cells are normal human dermal fibroblasts). 423 HTEEW69 Immune Highly preferred indications include SK-N-MC TNF gb|AJ270944|HSA27094 immunological disorders such as described herein neuro- VCAM gb|A30922|A30922 under the heading “Immune Activity” and/or blastoma “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving the central nervous system). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving the central nervous sytem). (The SK-N-MC neuroblastoma cell line is a cell line derived from human brain tissue and is available through the ATCC as cell line number HTB-10). 423 HTEEW69 Immune Highly preferred indications include THP1 CD25 gb|X03137|HSIL2RG7 immunological disorders such as described herein CD40 gb|AJ300189|HSA30018 under the heading “Immune Activity” and/or GATA3 gb|X55037|HSGATA3 “Blood-Related Disorders” (particularly LTBR gb|AK027080|AK027080 including, but not limited to, immune disorders Rag1 gb|M29474|HUMRAG1 involving monocytes). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving monocytes). (The THP1 cell line is a human monocyte cell line available through the ATCC as cell line number TIB-202). 423 HTEEW69 Immune Highly preferred indications include U937 IL1B gb|X02532|HSIL1BR immunological disorders such as described herein TNF gb|AJ270944|HSA27094 under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving monocytes). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving monocytes). (The U937 cell line is a human monocyte cell line available through the ATCC; cell #CRL-1593.2). 878 HCEGG08 Immune Highly preferred indications include AOSMC CIS3 gb|AB006967|AB006967 immunological disorders such as described herein Granzyme B gb|J04071|HUMCSE under the heading “Immune Activity” and/or IL1B gb|X02532|HSIL1BR “Blood-Related Disorders” (particularly IL5 gb|X12705|HSBCDFIA including, but not limited to, immune disorders involving muscle tissues and the cardiovascular system (e.g. heart, lungs, circulatory system)). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving muscle tissue or the cardiovascular system). (AOSMC cells are human aortic smooth muscle cells). 878 HCEGG08 Immune Highly preferred indications include HEK293 ICAM gb|X06990|HSICAM1 immunological disorders such as described herein under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving epithelial cells or the renal system). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving epithelial cells or the renal system). (The 293 cell line is a human embryonal kidney epithelial cell line available through the ATCC as cell line number CRL-1573). 878 HCEGG08 Immune Highly preferred indications include HUVEC CCR7 gb|X84702|HSDNABLR2 immunological disorders such as described herein TNF gb|AJ270944|HSA27094 under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving endothelial cells). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving endothelial cells). (HUVEC cells are human umbilical vein endothelial cells). 878 HCEGG08 Immune Highly preferred indications include Jurkat GATA1 gb|X17254|HSERYF1 immunological disorders such as described herein Rag1 gb|M29474|HUMRAG1 under the heading “Immune Activity” and/or Rag2 gb|AY011962|AY011962 “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving T-cells). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving T-cells). (The Jurkat cell line is a human T lymphocyte cell line available through the ATCC; cell line #TIB-152). 878 HCEGG08 Immune Highly preferred indications include Liver ICAM gb|X06990|HSICAM1 immunological disorders such as described herein under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving cells of the hepatic system). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving cells of the hepatic system). 878 HCEGG08 Immune Highly preferred indications include NHDF HLA-c immunological disorders such as described herein under the heading “Immune Activity” and/or “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving the skin). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving the skin). (NHDF cells are normal human dermal fibroblasts). 878 HCEGG08 Immune Highly preferred indications include SK-N-MC HLA-c immunological disorders such as described herein neuro- VCAM gb|A30922|A30922 under the heading “Immune Activity” and/or blastoma “Blood-Related Disorders” (particularly including, but not limited to, immune disorders involving the central nervous system). Highly preferred embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving the central nervous sytem). (The SK-N-MC neuroblastoma cell line is a cell line derived from human brain tissue and is available through the ATCC as cell line number HTB-10). 878 HCEGG08 Immune Highly preferred indications include THP1 CCR3 gb|AB023887|AB023887 immunological disorders such as described herein CCR4 gb|AB023888|AB023888 under the heading “Immune Activity” and/or CTLA4 gb|AF316875|AF316875 “Blood-Related Disorders” (particularly Granzyme B gb|J04071|HUMCSE including, but not limited to, immune disorders Rag2 gb|AY011962|AY011962 involving monocytes). Highly preferred VCAM gb|A30922|A30922 embodiments of the invention include methods of preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving monocytes). (The THP1 cell line is a human monocyte cell line available through the ATCC as cell line number TIB-202). 878 HCEGG08 Immune Highly preferred indications include U937 CCR5 gb|AF161918|AF161918 immunological disorders such as described herein CCR7 gb|X84702|HSDNABLR2 under the heading “Immune Activity” and/or CD25 gb|X03137|HSIL2RG7 “Blood-Related Disorders” (particularly CD30 including, but not limited to, immune disorders CXCR3 gb|Z79783|HSCKRL2 involving monocytes). Highly preferred Rag1 gb|M29474|HUMRAG1 embodiments of the invention include methods of Rag2 gb|AY011962|AY011962 preventing, detecting, diagnosing, treating and/or ameliorating disorders of the immune system (particularly including, but not limited to, immune disorders involving monocytes). (The U937 cell line is a human monocyte cell line available through the ATCC as cell line number CRL-1593.2).

Table 2 further characterizes certain encoded polypeptides of the invention, by providing the results of comparisons to protein and protein family databases. The first column provides a unique clone identifier, “Clone ID NO:”, corresponding to a cDNA clone disclosed in Table 1A and/or Table 1B. The second column provides the unique contig identifier, “Contig ID:” which allows correlation with the information in Table 1B. The third column provides the sequence identifier, “SEQ ID NO:”, for the contig polynucleotide sequences. The fourth column provides the analysis method by which the homology/identity disclosed in the Table was determined. The fifth column provides a description of the PFAM/NR hit identified by each analysis. Column six provides the accession number of the PFAM/NR hit disclosed in the fifth column. Column seven, score/percent identity, provides a quality score or the percent identity, of the hit disclosed in column five. Comparisons were made between polypeptides encoded by polynucleotides of the invention and a non-redundant protein database (herein referred to as “NR”), or a database of protein families (herein referred to as “PFAM”), as described below.

The NR database, which comprises the NBRF PIR database, the NCBI GenPept database, and the SIB SwissProt and TrEMBL databases, was made non-redundant using the computer program nrdb2 (Warren Gish, Washington University in Saint Louis). Each of the polynucleotides shown in Table 1B, column 3 (e.g., SEQ ID NO:X or the ‘Query’ sequence) was used to search against the NR database. The computer program BLASTX was used to compare a 6-frame translation of the Query sequence to the NR database (for information about the BLASTX algorithm please see Altshul et al., J. Mol. Biol. 215:403-410 (1990), and Gish and States, Nat. Genet. 3:266-272 (1993). A description of the sequence that is most similar to the Query sequence (the highest scoring ‘Subject’) is shown in column five of Table 2 and the database accession number for that sequence is provided in column six. The highest scoring ‘Subject’ is reported in Table 2 if (a) the estimated probability that the match occurred by chance alone is less than 1.0e-07, and (b) the match was not to a known repetitive element. BLASTX returns alignments of short polypeptide segments of the Query and Subject sequences which share a high degree of similarity; these segments are known as High-Scoring Segment Pairs or HSPs. Table 2 reports the degree of similarity between the Query and the Subject for each HSP as a percent identity in Column 7. The percent identity is determined by dividing the number of exact matches between the two aligned sequences in the HSP, dividing by the number of Query amino acids in the HSP and multiplying by 100. The polynucleotides of SEQ ID NO:X which encode the polypeptide sequence that generates an HSP are delineated by columns 8 and 9 of Table 2.

The PFAM database, PFAM version 2.1, (Sonnhammer, Nucl. Acids Res., 26:320-322, 1998)) consists of a series of multiple sequence alignments; one alignment for each protein family. Each multiple sequence alignment is converted into a probability model called a Hidden Markov Model, or HMM, that represents the position-specific variation among the sequences that make up the multiple sequence alignment (see, e.g., Durbin, et al., Biological sequence analysis: probabilistic models of proteins and nucleic acids, Cambridge University Press, 1998 for the theory of HMMs). The program HMMER version 1.8 (Sean Eddy, Washington University in Saint Louis) was used to compare the predicted protein sequence for each Query sequence (SEQ ID NO:Y in Table 1B.1) to each of the HMMs derived from PFAM version 2.1. A HMM derived from PFAM version 2.1 was said to be a significant match to a polypeptide of the invention if the score returned by HMMER 1.8 was greater than 0.8 times the HMMER 1.8 score obtained with the most distantly related known member of that protein family. The description of the PFAM family which shares a significant match with a polypeptide of the invention is listed in column 5 of Table 2, and the database accession number of the PFAM hit is provided in column 6. Column 7 provides the score returned by HMMER version 1.8 for the alignment. Columns 8 and 9 delineate the polynucleotides of SEQ ID NO:X which encode the polypeptide sequence which show a significant match to a PFAM protein family.

As mentioned, columns 8 and 9 in Table 2, “NT From” and “NT To”, delineate the polynucleotides of “SEQ ID NO:X” that encode a polypeptide having a significant match to the PFAM/NR database as disclosed in the fifth column. In one embodiment, the invention provides a protein comprising, or alternatively consisting of, a polypeptide encoded by the polynucleotides of SEQ ID NO:X delineated in columns 8 and 9 of Table 2. Also provided are polynucleotides encoding such proteins, and the complementary strand thereto.

The nucleotide sequence SEQ ID NO:X and the translated SEQ ID NO:Y are sufficiently accurate and otherwise suitable for a variety of uses well known in the alt and described further below. For instance, the nucleotide sequences of SEQ ID NO:X are useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ID NO:X or the cDNA contained in ATCC Deposit No:Z. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling immediate applications in chromosome mapping, linkage analysis, tissue identification and/or typing, and a variety of forensic and diagnostic methods of the invention. Similarly, polypeptides identified from SEQ ID NO:Y may be used to generate antibodies which bind specifically to these polypeptides, or fragments thereof, and/or to the polypeptides encoded by the cDNA clones identified in, for example, Table 1A and/or 1B.

Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence. In these cases, the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).

Accordingly, for those applications requiring precision in the nucleotide sequence or the amino acid sequence, the present invention provides not only the generated nucleotide sequence identified as SEQ ID NO:X, and a predicted translated amino acid sequence identified as SEQ ID NO:Y, but also a sample of plasmid DNA containing cDNA ATCC Deposit No:Z (e.g., as set forth in columns 2 and 3 of Table 1A and/or as set forth, for example, in Table 1B, 6, and 7). The nucleotide sequence of each deposited clone can readily be determined by sequencing the deposited clone in accordance with known methods. Further, techniques known in the art can be used to verify the nucleotide sequences of SEQ ID NO:X.

The predicted amino acid sequence can then be verified from such deposits. Moreover, the amino acid sequence of the protein encoded by a particular clone can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human cDNA, collecting the protein, and determining its sequence. TABLE 2 SEQ PFam/NR Score/ cDNA Contig ID Analysis Accession Percent NT Clone ID ID: NO: X Method PFam/NR Description Number Identity From NT To H6BSF56 762968 11 HMMER PFAM: Zinc-binding dehydrogenases PF00107 35.6 176 415 2.1.1 WUblastx.64 (Q9BV79) SIMILAR TO CGI-63 Q9BV79 100% 25 42 PROTEIN. 92% 53 427 H6EDM64 841331 12 WUblastx.64 (Q9UID3) ANG2. Q9UID3 90% 928 2451 36% 203 310 36% 931 1038 95% 191 871 H6EEC72 889401 13 WUblastx.64 hypothetical protein DKFZp434L061.1 - pir|T43456|T43456 80% 1484 1203 human 41% 1277 1080 35% 973 845 34% 659 549 57% 991 365 HACBJ56 847112 15 WUblastx.64 (Q9D2Q2) 2310079F23RIK Q9D2Q2 65% 98 286 PROTEIN. HACBS22 847113 16 WUblastx.64 (O60266) ADENYLATE CYCLASE CYA3_HUMAN 89% 6 416 TYPE III (EC 4.6.1.1) (ADENYLATE 25% 1547 2299 18% 917 1111 93% 416 2449 HADMB15 847116 19 WUblastx.64 (Q9BVH1) SIMILAR TO DLXIN-1. Q9BVH1 100% 8 109 HAGEG10 823543 22 WUblastx.64 (Q9NWT5) CDNA FLJ20618 FIS, Q9NWT5 100% 1237 1377 CLONE KAT05049. 96% 1 156 HAGFS57 847120 24 WUblastx.64 (Q9Y485) X-LIKE 1 PROTEIN. Q9Y485 58% 9 872 HAGHN57 773286 25 WUblastx.64 (O60416) WUGSC: H_RG276O03.2 O60416 98% 65 1444 PROTEIN. HAHEA15 847013 26 WUblastx.64 (Q9NWD5) HYPOTHETICAL 31.4 KDA Q9NWD5 76% 455 832 PROTEIN. 99% 30 560 HAJAA47 534670 27 WUblastx.64 (Q9NZA3) CDA14. Q9NZA3 100% 17 157 HAJAY92 845601 28 WUblastx.64 (O00549) ORF2-LIKE PROTEIN O00549 53% 2226 2318 (FRAGMENT). 26% 769 915 38% 1653 1769 31% 1721 2242 HAJBV67 866415 29 WUblastx.64 (Q9HD45) TRANSMEMBRANE 9 T9S3_HUMAN 100% 13 126 SUPERFAMILY PROTEIN 93% 116 1681 MEMBER 3 PRECU HAOAG15 852204 31 HMMER PFAM: von Willebrand factor type A PF00092 180.1 506 1057 2.1.1 domain WUblastx.64 (O75578) INTEGRIN ALPHA-10 ITAG_HUMAN 90% 8 3463 PRECURSOR. HAQCE11 633730 33 WUblastx.64 (Q24333) ELASTIN LIKE PROTEIN Q24333 95% 61 132 (FRAGMENT). HATCB45 631172 35 WUblastx.64 (Q9D0I6) 2610014F08RIK PROTEIN. Q9D0I6 88% 490 645 HATCI03 580805 37 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 71% 906 688 CLONE COL03536. HBAGD86 838799 39 WUblastx.64 (Q14287) HYPOTHETICAL Q14287 37% 801 559 PROTEIN (FRAGMENT). HBDAB91 789532 41 WUblastx.64 (O00370) PUTATIVE P150. O00370 40% 587 513 34% 529 5 HBDAB91 864374 42 WUblastx.64 (O00370) PUTATIVE P150. O00370 40% 907 833 35% 849 307 HBGBC29 691473 43 WUblastx.64 (O60513) BETA-1,4- B4G4_HUMAN 61% 1 78 GALACTOSYLTRANSFERASE 4 98% 65 1021 (EC 2.4.1.—) (BET HBHAA05 603174 45 WUblastx.64 (Q9H387) PRO2550. Q9H387 71% 676 386 HBHAA81 846465 46 WUblastx.64 (Q9D1G3) 1110011D13RIK Q9D1G3 89% 1329 1502 PROTEIN. 79% 28 1329 HBIAC29 831751 48 WUblastx.64 (Q9D7J5) 2310005N01RIK Q9D7J5 78% 25 492 PROTEIN. 93% 883 927 HBJAB02 837309 50 WUblastx.64 (Q9NXT6) CDNA FLJ20062 FIS, Q9NXT6 70% 2 1210 CLONE COL01508. HBJCR46 815649 53 HMMER PFAM: WD domain, G-beta repeat PF00400 36.6 790 867 2.1.1 WUblastx.64 (Q9DC22) 1200006M05RIK Q9DC22 96% 207 611 PROTEIN. 73% 568 2763 HBJDS79 813588 54 WUblastx.64 (Q9CY11) 2510039O18RIK Q9CY11 92% 1119 1325 PROTEIN. 89% 1322 1519 93% 1032 1127 100% 1509 1532 66% 2 1075 HBJEL16 847030 56 WUblastx.64 (O95297) PROTEIN ZERO O95297 98% 285 491 RELATED PROTEIN. HBJIG20 866159 58 HMMER PFAM: Cytochrome c oxidase subunit PF00510 162.6 321 551 2.1.1 III WUblastx.64 (BAA77671) Cytochrome c oxidase BAA77671 81% 9 617 subunit 3 (Fragment HBJKD16 853358 59 WUblastx.64 (Q9NXS4) CDNA FLJ20080 FIS, Q9NXS4 91% 8 1528 CLONE COL03184. HBMBM96 561935 60 WUblastx.64 (Q9H387) PRO2550. Q9H387 69% 661 494 67% 794 639 HBMTX26 695704 63 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 46% 964 608 PROTEIN (FRAGMENT). 61% 272 156 66% 136 101 54% 611 507 58% 546 292 HBMTY48 637521 64 WUblastx.64 (Q9H5N9) CDNA: FLJ23235 FIS, Q9H5N9 94% 54 941 CLONE CAS04980. HBMUH74 866160 65 WUblastx.64 (Q9NVW8) CDNA FLJ10462 FIS, Q9NVW8 100% 11 427 CLONE NT2RP1001494, WEAKLY SIMILAR TO MAL HBMWE61 778066 66 WUblastx.64 (Q9BX88) MAGPHININ DELTA. Q9BX88 100% 302 520 95% 869 1009 HBNAX40 834801 67 WUblastx.64 (Q9H2K2) TANKYRASE-LIKE Q9H2K2 100% 1 201 PROTEIN (TANKYRASE 2). 100% 221 481 HBQAB79 810542 69 WUblastx.64 (Q9UQ32) AD 3 (FRAGMENT). Q9UQ32 82% 323 204 HBSAK32 856387 71 WUblastx.64 (Q9H1Q7) BA12M19.1.3 (NOVEL Q9H1Q7 100% 239 412 PROTEIN). 100% 95 172 HBXCM66 639039 72 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 65% 988 809 CLONE COL04765. 77% 836 690 HBXCX15 637542 73 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 41% 726 827 PROTEIN. 52% 578 730 HCDCY76 837972 74 WUblastx.64 frizzled protein 4 - human pir|JC7127|JC7127 100% 1039 527 30% 994 785 79% 567 37 HCE1G78 761204 76 HMMER PFAM: Inositol polyphosphate PF00783 277.3 77 775 2.1.1 phosphatase family, catalytic domain WUblastx.64 (Q9UDT9) WUGSC: H_DJ412A9.2 Q9UDT9 72% 8 1549 PROTEIN (FRAGMENT). 95% 8 67 HCE2H52 847007 77 WUblastx.64 probable transposase - human pir|S72481|S72481 60% 564 758 transposon MER37 77% 430 537 75% 754 1251 HCE3B04 831151 78 WUblastx.64 (O43466) HYPOTHETICAL 31.3 KDA O43466 98% 836 1003 PROTEIN (FRAGMENT). 45% 217 972 HCEDR26 771144 80 WUblastx.64 (Q9H919) CDNA FLJ13078 FIS, Q9H919 66% 1157 1095 CLONE NT2RP3002002. 66% 1345 1184 HCEEQ25 531784 82 WUblastx.64 (P78349) SODIUM CHANNEL 2. P78349 95% 311 433 93% 433 480 100% 658 714 HCEEU18 688041 83 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 49% 186 10 PRODUCT. 56% 1223 933 HCEFZ82 831745 84 WUblastx.64 (Q9BV23) SIMILAR TO LIPASE Q9BV23 95% 594 782 PROTEIN. 100% 17 604 HCFLN88 610000 86 WUblastx.64 (Q9BQE9) SIMILAR TO B-CELL Q9BQE9 87% 278 475 CLL/LYMPHOMA 7B (UNKNOWN) (PROTEIN FOR MGC HCFLT90 788578 87 WUblastx.64 (Q9CVC2) 2210013O21RIK Q9CVC2 53% 612 445 PROTEIN (FRAGMENT). 70% 850 671 HCHAB84 834326 88 WUblastx.64 (Q9BRV3) STROMAL CELL Q9BRV3 89% 82 744 PROTEIN. HCNSD29 862314 91 WUblastx.64 (O75400) HUNTINGTIN- O75400 82% 628 1605 INTERACTING PROTEIN 78% 337 489 HYPA/FBP11 (FRAGMENT). HCRAY10 695709 96 WUblastx.64 (AAH08671) Similar to RIKEN cDNA AAH08671 77% 72 440 5530601I19 gene. HCRBF72 828945 97 WUblastx.64 (Q9UI95) MITOTIC SPINDLE MD22_HUMAN 94% 191 823 ASSEMBLY CHECKPOINT PROTEIN MAD2B HCUCF89 637986 100 WUblastx.64 (Q9P147) PRO2822. Q9P147 100% 421 398 82% 494 426 HCUCK44 790277 101 WUblastx.64 hypothetical protein DKFZp564J157.1 - pir|T34520|T34520 100% 29 157 human (fragment) 100% 377 403 HDPDI72 897277 109 WUblastx.64 adult-specific brush border protein - pir|C45665|C45665 64% 180 230 rabbit 83% 11 100 HDPDJ58 587265 110 WUblastx.64 hypothetical protein pir|T42691|T42691 100% 307 609 DKFZp434D2328.1 - human 87% 621 785 (fragment) 36% 101 313 36% 307 606 36% 188 316 42% 89 172 27% 23 307 85% 14 307 37% 134 307 35% 101 307 35% 89 274 32% 137 307 36% 325 594 34% 543 671 38% 307 606 37% 322 585 28% 307 606 29% 358 606 33% 337 594 41% 487 594 37% 92 307 36% 352 606 32% 89 316 32% 340 594 30% 83 316 35% 454 606 31% 654 785 41% 624 779 31% 624 794 40% 630 785 34% 624 776 33% 630 785 34% 83 307 35% 92 286 34% 1229 1402 36% 1265 1411 36% 660 785 32% 624 779 39% 645 773 35% 645 785 35% 645 785 32% 785 1393 34% 280 606 36% 259 594 32% 259 591 32% 319 591 100% 1405 1464 33% 322 510 33% 633 881 29% 656 1327 25% 902 1402 30% 848 1402 30% 1040 1420 36% 89 307 39% 827 1042 41% 962 1117 27% 803 1402 33% 131 307 25% 848 1144 27% 89 307 37% 125 307 30% 854 1447 76% 746 1450 HDPFF10 853513 111 HMMER PFAM: Leucine Rich Repeat PF00560 65.1 729 800 2.1.1 WUblastx.64 garp precursor - human pir|S42799|S42799 38% 285 965 42% 1153 1593 32% 1306 1641 29% 1147 1446 31% 1614 1898 26% 1159 1512 30% 1174 1536 33% 1306 1632 30% 1174 1494 31% 1162 1539 33% 1183 1500 28% 468 893 27% 246 965 37% 1629 2111 HDPFU43 790189 112 WUblastx.64 (AAH01057) Tyrosylprotein AAH01057 100% 360 1349 sulfotransferase 2. 58% 220 348 HDPGE24 801947 114 WUblastx.64 (Q9P195) PRO1722. Q9P195 65% 1413 1291 43% 1388 1278 77% 2528 2394 47% 2182 2078 75% 1774 1751 62% 2604 2557 68% 1301 1167 HDPIU94 813352 115 WUblastx.64 (Q9BVF7) SIMILAR TO Q9BVF7 99% 63 1703 HYPOTHETICAL PROTEIN FLJ10422. HDPOC24 777493 116 WUblastx.64 (Q9H8K1) CDNA FLJ13518 FIS, Q9H8K1 100% 62 208 CLONE PLACE1005799. HDPOL37 745377 117 WUblastx.64 (AAK40301) TRH4. AAK40301 70% 502 323 60% 1325 483 HDPPQ30 684292 120 WUblastx.64 (Q9H387) PRO2550. Q9H387 51% 807 727 79% 1042 815 HDPPW82 778405 121 WUblastx.64 hypothetical protein UL126 - human pir|S09875|S09875 94% 6 116 cytomegalovirus (strain AD169) HDQHM36 852328 123 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 69% 1129 1257 PRODUCT. 50% 965 1153 HDTAU35 838139 124 WUblastx.64 (Q9T9V8) NADH Q9T9V8 87% 56 175 DEHYDROGENASE SUBUNIT 3. 83% 305 340 HDTAV54 801898 125 WUblastx.64 (AAH01231) Glutathione S-transferase AAH01231 100% 13 303 subunit 13 hom HDTGW48 827285 127 WUblastx.64 (Q9P1W8) SIRP-B2. Q9P1W8 100% 783 1100 79% 1359 1757 HE2CA60 888705 130 WUblastx.64 (O95232) OKADAIC ACID- OA48_HUMAN 98% 1098 1265 INDUCIBLE PHOSPHOPROTEIN OA48-18. HE2HC60 753265 133 WUblastx.64 (Q9NVC4) CDNA FLJ10814 FIS, Q9NVC4 88% 125 1300 CLONE NT2RP4000984. HE6CS65 762960 136 WUblastx.64 (Q9H7C6) CDNA: FLJ21047 FIS, Q9H7C6 98% 938 1378 CLONE CAS00253. HE6DO92 562767 137 WUblastx.64 gag polyprotein - human endogenous pir|A46312|A46312 63% 623 895 virus S71 80% 19 633 HE6EY13 847058 138 WUblastx.64 (O95476) HYPOTHETICAL 28.3 KDA O95476 92% 5 472 PROTEIN. HE6FU11 827236 139 HMMER PFAM: von Willebrand factor type A PF00092 184.7 244 771 2.1.1 domain WUblastx.64 (O95460) MATRILIN-4 MTN4_HUMAN 77% 145 789 PRECURSOR. 45% 782 907 41% 791 925 50% 794 907 38% 863 1498 33% 190 741 98% 782 1642 HE8FC45 843781 141 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 50% 1285 1172 CLONE KAIA0536. 57% 1824 1663 75% 1672 1553 HE8FC45 845672 142 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 50% 1285 1172 CLONE KAIA0536. 57% 1824 1663 75% 1672 1553 HE8FD92 856544 145 WUblastx.64 (Q9UJI9) HYPOTHETICAL 105.9 KDA Q9UJI9 84% 419 1414 PROTEIN. 71% 2 1060 76% 419 1345 61% 2 613 45% 203 1060 40% 449 1345 52% 2 328 33% 605 1345 46% 47 328 HE8FD92 869847 146 WUblastx.64 (Q9UJI9) HYPOTHETICAL 105.9 KDA Q9UJI9 74% 4 609 PROTEIN. 59% 4 540 50% 1 255 63% 346 540 63% 346 540 60% 346 540 49% 1 255 48% 1 255 53% 346 540 41% 4 255 33% 40 255 HE8FD92 901142 147 WUblastx.64 (Q9UJI9) HYPOTHETICAL 105.9 KDA Q9UJI9 76% 31 480 PROTEIN. 56% 31 411 67% 217 408 67% 217 408 63% 217 411 63% 217 411 60% 217 411 53% 217 411 59% 31 126 56% 31 126 56% 31 126 56% 31 126 39% 58 126 HE8SG96 862016 148 WUblastx.64 (Q9P195) PRO1722. Q9P195 58% 1997 1845 63% 1854 1687 HE8TY46 899528 149 WUblastx.64 (BAB55144) CDNA FLJ14576 fis, BAB55144 95% 318 938 clone NT2RM4001092, w HE9CY05 834826 150 WUblastx.64 (Q9CX63) 6030468B19RIK Q9CX63 48% 434 742 PROTEIN. 57% 55 426 HE9EA10 827796 151 WUblastx.64 laminin alpha-1 chain precursor - pir|S14458|S14458 99% 761 1891 human 27% 878 1840 25% 1142 1876 HEBFR46 847064 157 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 80% 1111 1022 CLONE KAIA0536. 84% 1265 1110 HEBGE07 798096 158 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 79% 1851 1720 CLONE KAIA0536. HELAT35 693175 160 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 72% 2092 1802 CLONE COL04765. HELBU54 637624 161 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 59% 1255 1031 CLONE COL04765. HEMEY47 834491 164 WUblastx.64 (Q9H387) PRO2550. Q9H387 68% 513 587 74% 578 838 HEPBA14 855935 166 WUblastx.64 (Q9BTY9) UNKNOWN (PROTEIN Q9BTY9 87% 423 515 FOR IMAGE: 2823490) 71% 15 77 (FRAGMENT). 92% 85 426 HEQAH80 701984 167 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 60% 818 1045 PROTEIN. HEQBF89 786205 168 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 64% 793 638 CLONE COL04765. 64% 647 489 HETCI16 844543 169 WUblastx.64 (Q9P0V3) BOG25. Q9P0V3 99% 3 356 HETDW58 790557 170 WUblastx.64 unidentified 27.6K protein, spliced pir|JC7586|JC7586 95% 324 1058 form A - human HFCFE20 701985 175 WUblastx.64 (Q9CSE5) EUKARYOTIC Q9CSE5 89% 438 581 TRANSLATION INITIATION 54% 1083 1187 FACTOR 3 (FRAGMENT). HFEAY59 658685 176 WUblastx.64 (Q9Z320) C29. Q9Z320 67% 50 1153 HFGAJ16 580824 177 WUblastx.64 CDM protein - human pir|S44279|S44279 97% 263 403 HFIJA29 839206 179 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 46% 889 806 59% 1026 880 HFIJA68 847074 180 WUblastx.64 (Q9UHE8) SIX TRANSMEMBRANE STEA_HUMAN 89% 13 399 EPITHELIAL ANTIGEN OF PROSTATE. HFKES05 827572 181 WUblastx.64 (BAB55088) CDNA FLJ14496 fis, BAB55088 85% 84 314 clone NT2RM1000035. 94% 367 1722 HFKEU12 634006 182 WUblastx.64 hypothetical protein 3 - rat pir|S21347|S21347 52% 695 745 50% 757 933 40% 774 1007 54% 387 692 HFPDS07 821646 185 WUblastx.64 (O94925) GLUTAMINASE, KIDNEY GLSK_HUMAN 78% 343 513 ISOFORM, MITOCHONDRIAL 74% 2 436 PRECURS HFVGK35 731868 189 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 65% 832 608 PROTEIN. HFVHW43 570948 190 WUblastx.64 (Q9BGX4) HYPOTHETICAL 13.8 KDA Q9BGX4 69% 1209 1093 PROTEIN. HFXAV37 626595 191 WUblastx.64 (O60448) NEURONAL THREAD O60448 33% 583 461 PROTEIN AD7C-NTP. 68% 473 333 60% 1240 1166 47% 607 539 45% 1454 1275 56% 1295 1173 59% 1287 1222 64% 607 407 69% 607 461 57% 1285 1169 40% 1467 1402 48% 1321 1232 37% 558 346 78% 402 361 68% 591 406 31% 549 355 60% 398 354 36% 1321 1232 41% 1364 1275 46% 1187 1110 47% 1453 1187 HFXBT66 580831 193 WUblastx.64 (Q9H387) PRO2550. Q9H387 73% 739 807 58% 809 907 62% 564 764 HGBER72 826710 195 WUblastx.64 (Q9H387) PRO2550. Q9H387 71% 1061 969 78% 1104 1063 77% 1237 1103 HGBHP91 693011 198 WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 52% 541 491 human 44% 537 34 HGCAC19 851527 199 WUblastx.64 (Q9UIE9) WUGSC: H_DJ0687K01.2 Q9UIE9 34% 984 1124 PROTEIN. 31% 1546 1863 96% 361 1047 24% 993 1124 22% 984 1124 40% 1002 1124 27% 984 1124 27% 981 1124 25% 984 1088 31% 984 1124 23% 984 1124 29% 984 1124 29% 984 1124 32% 1023 1124 26% 586 801 95% 2017 2712 HGCAC19 801999 200 WUblastx.64 (Q9H6A1) CDNA: FLJ22454 FIS, Q9H6A1 34% 984 1124 CLONE HRC09703 (FRAGMENT). 100% 184 210 31% 984 1124 29% 984 1124 40% 1002 1124 27% 984 1124 29% 984 1124 27% 981 1124 21% 984 1121 24% 993 1124 23% 984 1124 96% 316 1056 HGCAC19 842540 201 WUblastx.64 (Q9H6A1) CDNA: FLJ22454 FIS, Q9H6A1 34% 982 1122 CLONE HRC09703 (FRAGMENT). 100% 182 208 31% 982 1122 29% 982 1122 40% 1000 1122 27% 982 1122 29% 982 1122 27% 979 1122 21% 982 1119 24% 991 1122 23% 982 1122 96% 314 1045 HHEAK45 765278 202 WUblastx.64 (Q9NPB0) DJ202I21.1 (NOVEL Q9NPB0 68% 1949 1458 PROTEIN) (CDNA FLJ11101 FIS, CLONE PLACE10 HHEOW19 886174 204 WUblastx.64 (O18973) RAB5 GDP/GTP O18973 77% 417 623 EXCHANGE FACTOR, RABEX5. 91% 611 715 56% 166 378 92% 129 167 HHFFF87 778071 205 WUblastx.64 coatomer zeta chain - bovine pir|A49465|A49465 100% 50 145 HHFFL34 753230 206 WUblastx.64 (BAB55306) CDNA FLJ14793 fis, BAB55306 100% 9 710 clone NT2RP4001174, w HHFFS40 824059 207 WUblastx.64 (Q9H4A6) GOLGI PROTEIN. Q9H4A6 100% 3 251 HHGDT26 658692 209 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 69% 1580 1290 CLONE COL04765. HHSBI65 801910 213 WUblastx.64 (Q9H5W9) CDNA: FLJ22888 FIS, Q9H5W9 100% 270 407 CLONE KAT03934. 94% 479 1300 HHSDI53 862028 214 WUblastx.64 (Q9H387) PRO2550. Q9H387 70% 1108 935 71% 1241 1107 75% 1276 1241 HILCA24 782450 217 WUblastx.64 (Q9NUU6) CDNA FLJ11127 FIS, Q9NUU6 73% 103 159 CLONE PLACE1006225. 100% 168 1169 HILCA24 869856 218 WUblastx.64 (Q9NUU6) CDNA FLJ11127 FIS, Q9NUU6 95% 104 1171 CLONE PLACE1006225. HISAT67 843549 219 WUblastx.64 (Q9UH94) PROLACTIN Q9UH94 88% 219 797 REGULATORY ELEMENT- 91% 788 1447 BINDING PROTEIN (PROLACTIN REGU HJBCU75 638329 220 WUblastx.64 (O45030) STRABISMUS. O45030 44% 199 426 52% 464 964 HJMAA03 824062 221 WUblastx.64 (Q9N032) UNNAMED PROTEIN Q9N032 71% 415 528 PRODUCT. HJMAV41 862029 222 WUblastx.64 brain-specific membrane anchor pir|JC7110|JC7110 100% 14 475 protein - human HJMAY90 793678 223 WUblastx.64 (Q9DC16) 1200007D18RIK Q9DC16 77% 100 312 PROTEIN (RIKEN CDNA 98% 315 968 1200007D18 GENE). HJPBE39 801960 224 WUblastx.64 (Q9CUS4) 4833420K19RIK Q9CUS4 33% 1 621 PROTEIN (FRAGMENT). 74% 213 1007 HJPCH08 840365 226 WUblastx.64 (O95235) RABKINESIN-6 (RAB6- RB6K_HUMAN 93% 9 596 INTERACTING KINESIN-LIKE PROTEI HKABU43 838573 227 WUblastx.64 (AAH03633) Translocase of outer AAH03633 100% 33 62 mitochondrial membr 92% 26 1597 HKACI79 853361 228 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 72% 886 1104 PROTEIN. HKAFF50 790192 229 WUblastx.64 (Q9P1G7) PRO1777. Q9P1G7 99% 1753 1424 HKGBF25 738797 230 WUblastx.64 (Q9HBS7) HYPOTHETICAL 14.2 KDA Q9HBS7 71% 1708 1688 PROTEIN. 56% 1956 1708 HKMLK03 734213 232 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 50% 981 832 PRODUCT. 73% 856 731 HLDQU79 740755 237 WUblastx.64 (O75477) KE04P. O75477 100% 105 1142 HLDQU79 837599 3099 blastx.2 KE04P. sp|O75477|O75477 99% 81 1118 HLDRT09 830544 238 WUblastx.64 (Q9HAQ7) ATP-BINDING Q9HAQ7 86% 2 469 CASSETTE HALF-TRANSPORTER. HLHAP05 638476 239 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 55% 1553 1500 CLONE MAMMA1000472. 72% 1650 1585 77% 1807 1646 HLIBO72 883431 241 WUblastx.64 (AAH07829) Similar to hypothetical AAH07829 100% 65 547 protein AF140225 HLICE88 840321 242 WUblastx.64 fibrinogen gamma-A chain precursor pir|A90470|FGHUG 89% 3 584 [validated] - human HLMBW89 701996 245 WUblastx.64 (AAH07983) Unknown (protein for AAH07983 85% 390 247 MGC: 16279). HLMGP50 647603 246 WUblastx.64 (Q9GMI7) HYPOTHETICAL 9.0 KDA Q9GMI7 61% 765 709 PROTEIN. 72% 935 807 HLQAS12 886180 249 WUblastx.64 (Q9XTA8) LECTIN-LIKE Q9XTA8 71% 690 842 OXIDIZED LDL RECEPTOR. 52% 364 711 HLQCL64 864966 250 HMMER PFAM: Major intrinsic protein PF00230 87.3 87 449 2.1.1 WUblastx.64 aquaporin 9 - human pir|JC5973|JC5973 98% 18 548 HLQCX36 584786 251 WUblastx.64 (Q9UI59) PRO0478 PROTEIN. Q9UI59 87% 1100 1216 HLWDB73 838453 258 WUblastx.64 (Q9H7D7) CDNA: FLJ21016 FIS, Q9H7D7 100% 660 872 CLONE CAE05735. 98% 1 657 HLYGB19 838083 261 WUblastx.64 (Q9H0Q1) HYPOTHETICAL 12.3 KDA Q9H0Q1 97% 204 518 PROTEIN. HLYGY91 658703 263 WUblastx.64 (Q9H8N0) CDNA FLJ13386 FIS, Q9H8N0 94% 221 391 CLONE PLACE1001104, WEAKLY SIMILAR TO MYO HMCAZ04 839783 264 WUblastx.64 (Q9Y6N5) HYPOTHETICAL 50.0 KDA Q9Y6N5 100% 106 1455 PROTEIN. HMCAZ04 858210 265 WUblastx.64 (Q9Y6N5) HYPOTHETICAL 50.0 KDA Q9Y6N5 100% 106 1455 PROTEIN. HMCAZ04 867910 266 WUblastx.64 (Q9Y6N5) HYPOTHETICAL 50.0 KDA Q9Y6N5 100% 106 1455 PROTEIN. HMCAZ04 887445 267 WUblastx.64 (Q9Y6N5) HYPOTHETICAL 50.0 KDA Q9Y6N5 100% 107 1456 PROTEIN. HMCAZ04 668249 268 WUblastx.64 (Q9UQM8) CGI-44 PROTEIN. Q9UQM8 100% 9 1055 HMDAB29 584789 270 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 72% 1186 890 CLONE KAT08285. HMEBB82 783077 272 WUblastx.64 (Q9NSE4) MITOCHONDRIAL Q9NSE4 99% 2 2206 ISOLEUCINE TRNA SYNTHETASE (FRAGMENT). HMEDE24 837027 273 WUblastx.64 (Q9BVH9) SIMILAR TO GLUCOSE Q9BVH9 94% 188 1159 REGULATED PROTEIN, 58 KDA. 42% 101 742 HMEDI90 840077 274 WUblastx.64 (Q9HBA3) RAB3 INTERACTING Q9HBA3 100% 81 794 PROTEIN VARIANT 4 (FRAGMENT). HMELM75 587307 275 WUblastx.64 (Q9NVW5) HYPOTHETICAL 31.3 KDA Q9NVW5 100% 137 391 PROTEIN. HMICP65 847403 279 WUblastx.64 (Q9HAU9) GUANINE Q9HAU9 99% 8 892 NUCLEOTIDE BINDING PROTEIN 22% 269 943 BETA SUBUNIT 5L. HMSBE04 709672 281 WUblastx.64 (Q9H5V8) CDNA: FLJ22969 FIS, Q9H5V8 85% 182 3 CLONE KAT10759. HMSCL38 801919 282 WUblastx.64 (Q9P195) PRO1722. Q9P195 64% 1272 1460 72% 2918 2844 64% 2851 2759 76% 2769 2653 HMSCR69 843059 283 HMMER PFAM: Zinc finger present in PF00569 48.2 113 250 2.1.1 dystrophin, CBP/p300 WUblastx.64 (Q9BWK2) POTASSIUM CHANNEL Q9BWK2 78% 107 1231 MODULATORY FACTOR. HMSHC86 840402 284 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 70% 1724 1674 PRODUCT. 67% 1674 1420 HMSHU20 847410 285 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 47% 1722 1453 CLONE COL04765. HMTAB77 847411 287 WUblastx.64 (P43243) MATRIN 3. MAT3_HUMAN 95% 630 1385 64% 287 628 22% 2002 2175 98% 3255 3428 31% 2041 2190 22% 2047 2181 23% 2584 2763 75% 2440 2760 27% 2596 2709 35% 1705 1797 35% 3312 3404 91% 1384 2328 HMUAE26 747403 288 WUblastx.64 (Q9P2R4) SEVEN Q9P2R4 89% 153 575 TRANSMEMBRANE DOMAIN 86% 577 1272 ORPHAN RECEPTOR. HMUAN45 833072 289 WUblastx.64 (BAB55441) CDNA FLJ14993 fis, BAB55441 70% 684 1238 clone Y79AA1001874, w 65% 239 955 100% 1247 1516 HMVBC31 825598 290 WUblastx.64 (O60725) PROTEIN-S ICMT_HUMAN 80% 747 938 ISOPRENYLCYSTEINE O- 87% 121 789 METHYLTRANSFERASE (E HMVDU15 801969 291 WUblastx.64 (Q9BTJ2) SIMILAR TO CGI-30 Q9BTJ2 100% 75 917 PROTEIN. HMWBL03 822861 292 WUblastx.64 (Q9BWT1) C-MYC TARGET JP1. Q9BWT1 85% 137 1240 HMWJF53 758158 293 WUblastx.64 (Q9GZU7) NUCLEAR LIM Q9GZU7 91% 3 170 INTERACTOR-INTERACTING 100% 154 720 FACTOR. HNEAK81 722235 294 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 56% 770 1087 PRODUCT. HNECL22 799541 295 WUblastx.64 (Q9P0J2) MITOCHONDRIAL Q9P0J2 94% 1771 2331 SOLUTE CARRIER. HNEDH88 815675 297 WUblastx.64 (Q9GML5) HYPOTHETICAL 8.0 KDA Q9GML5 56% 1706 1849 PROTEIN. HNFAC50 815676 298 WUblastx.64 (Q9H286) SEROLOGICALLY Q9H286 100% 425 282 DEFINED BREAST CANCER ANTIGEN NY-BR-20 (FRAGME HNFHF34 722237 300 WUblastx.64 (Q9NZX0) HSPC068. Q9NZX0 100% 9 431 34% 9 404 35% 3 407 33% 9 407 32% 129 422 HNGAK51 603910 301 WUblastx.64 (O60448) NEURONAL THREAD O60448 61% 563 601 PROTEIN AD7C-NTP. 67% 733 915 65% 702 878 74% 714 914 HNGAM58 688114 302 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 71% 1020 1061 CLONE COL04765. 85% 1081 1143 53% 818 1003 HNGGP65 597449 310 WUblastx.64 (Q9GMU5) HYPOTHETICAL 14.1 KDA Q9GMU5 31% 69 302 PROTEIN. 47% 398 541 HNGJB41 852178 313 WUblastx.64 probable oxysterol-binding protein pir|T02435|T02435 100% 128 9 DJ430N08.1 - human (fragment) HNHFE71 834487 320 WUblastx.64 hypothetical protein pir|T47135|T47135 67% 822 583 DKFZp761L0812.1 - human (fragment) HNHGK22 597451 321 WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 41% 483 37 human 41% 333 10 50% 733 485 HNHHB10 634589 322 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 70% 658 608 FOR MGC: 5149). 73% 845 711 73% 717 661 HNTBT17 855957 324 WUblastx.64 (Q9NZF3) BM-001. Q9NZF3 45% 818 1342 61% 729 947 84% 556 774 HOACG07 792928 328 WUblastx.64 (Q9GZN8) DJ1009E24.3 (A NOVEL Q9GZN8 99% 183 704 PROTEIN) (CDNA FLJ14158 FIS, CLONE NT2R HODBV05 825283 331 WUblastx.64 (Q13878) 94 KDA B-RAF PROTEIN Q13878 100% 566 661 (FRAGMENT). HODCZ32 836069 332 WUblastx.64 (Q9NSI6) WD-REPEAT PROTEIN 9 WDR9_HUMAN 86% 8 331 (FRAGMENT). HOEBK60 789396 333 WUblastx.64 (Q9H916) CDNA FLJ13081 FIS, Q9H916 98% 132 1916 CLONE NT2RP3002033. 100% 14 109 88% 106 159 HOFAA78 836646 334 WUblastx.64 (Q9NXS2) CDNA FLJ20084 FIS, Q9NXS2 90% 529 792 CLONE COL03526. 50% 9 80 88% 29 529 HOFNB74 762821 335 WUblastx.64 (Q99JH1) HYPOTHETICAL 17.7 KDA Q99JH1 72% 44 187 PROTEIN. 97% 199 471 HORBV76 839270 339 WUblastx.64 (Q9Y2B2) Q9Y2B2 91% 30 761 PHOSPHATIDYLINOSITOL GLYCAN, CLASS L (EC 3.5.—.—) (PIG-L PRO HOSDO75 862049 340 WUblastx.64 (Q9D099) 1110057L18RIK Q9D099 89% 11 202 PROTEIN. 88% 259 630 HOSEC25 688055 341 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 73% 530 631 PROTEIN. 65% 627 809 64% 1501 1451 56% 1440 1222 HOSEJ94 795132 343 WUblastx.64 (Q9GZY3) HT032 (PRK1- Q9GZY3 92% 363 986 ASSOCIATED PROTEIN AWP1) (PROTEIN ASSOCIATED WIT HOUCA21 655359 344 WUblastx.64 (Q9HBS7) HYPOTHETICAL 14.2 KDA Q9HBS7 78% 988 1110 PROTEIN. HOUDE92 580866 345 WUblastx.64 (Q9HBT2) HYPOTHETICAL 17.2 KDA Q9HBT2 96% 21 245 PROTEIN. HOUDR07 745404 346 WUblastx.64 (Q9HBV4) ANGIOPOIETIN-LIKE Q9HBV4 87% 170 1384 PROTEIN PP1158. HOUED72 858547 347 WUblastx.64 (Q9CRP8) RIBOSOMAL PROTEIN Q9CRP8 84% 676 774 L15 (FRAGMENT). 85% 110 682 HOUFS04 771564 348 WUblastx.64 (Q9VN45) CG12001 PROTEIN. Q9VN45 32% 1362 1982 39% 915 1106 26% 141 380 HOUHI25 888279 349 WUblastx.64 (O95003) WUGSC: H_DJ0593H12.2 O95003 94% 73 783 PROTEIN. HPCAL26 762822 352 WUblastx.64 (O95084) SERINE PROTEASE O95084 98% 398 640 (HYPOTHETICAL 43.0 KDA 76% 135 497 PROTEIN) (PROTEASE, S HPFBA54 635539 354 WUblastx.64 (Q9HBW6) NAG13. Q9HBW6 76% 795 733 73% 766 602 84% 602 393 86% 135 91 79% 394 128 HPFCI36 855966 355 WUblastx.64 (Q9NX47) CDNA FLJ20445 FIS, Q9NX47 100% 9 320 CLONE KAT05170. HPJBU43 862058 360 WUblastx.64 (Q9P1E1) PRO2221. Q9P1E1 54% 187 44 HPMCJ84 562779 363 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 74% 619 479 CLONE KAIA0536. 69% 759 613 HPMCV30 612870 364 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 76% 384 334 FOR MGC: 5149). 68% 590 399 HPQAX38 843592 366 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 74% 664 768 PROTEIN. 68% 543 674 HPQAX38 845752 367 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 74% 664 768 PROTEIN. 68% 543 674 HPRBH85 695752 370 WUblastx.64 (BAB55300) CDNA FLJ14784 fis, BAB55300 62% 2 616 clone NT2RP4000713. 86% 534 1085 HPRCA64 824074 371 WUblastx.64 (P55161) NCK-ASSOCIATED NCP1_RAT 100% 1021 1926 PROTEIN 1 (NAP 1) (P125NAP1) 85% 387 1019 (MEMBR 93% 11 481 HPRCD35 853551 372 WUblastx.64 hypothetical protein pir|T50629|T50629 100% 320 613 DKFZp762L1710.1 - human 57% 2 499 (fragment) HPTRM02 812879 373 WUblastx.64 (Q9UJU6) SRC HOMOLOGY 3 Q9UJU6 92% 332 940 DOMAIN-CONTAINING PROTEIN 97% 2 106 HIP-55 (DREBRIN F). 96% 98 190 HRAAD30 866187 376 WUblastx.64 (Q9H6V0) CDNA: FLJ21839 FIS, Q9H6V0 89% 23 1393 CLONE HEP01794. HRADA42 827302 377 WUblastx.64 hypothetical protein C11D2.4 - pir|T32961|T32961 48% 387 668 Caenorhabditis elegans 74% 668 931 HRADF49 866481 378 WUblastx.64 (Q9H6L1) CDNA: FLJ22169 FIS, Q9H6L1 90% 13 825 CLONE HRC00632. 84% 813 1379 75% 1291 1593 34% 1590 1685 HRADN25 800628 379 WUblastx.64 (Q9HB07) MYG1 PROTEIN. MYG1_HUMAN 96% 47 1174 HRDAI17 560720 381 WUblastx.64 (Q9NUM6) CDNA FLJ11267 FIS, Q9NUM6 59% 1305 1475 CLONE PLACE1009174. HRDDQ39 840405 382 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 53% 582 436 CLONE KAIA0536. 65% 775 578 HRDER22 688056 383 WUblastx.64 (Q9NW07) CDNA FLJ10390 FIS, Q9NW07 80% 9 248 CLONE NT2RM4000104, 100% 357 431 MODERATELY SIMILAR TO 39% 120 227 28% 15 203 38% 254 316 HRDEX93 816046 384 WUblastx.64 (Q9UBV8) PEFLIN. Q9UBV8 100% 313 864 HRDFK37 840381 385 WUblastx.64 (Q9P195) PRO1722. Q9P195 69% 536 652 40% 50 115 HRGBD54 828436 386 WUblastx.64 (O95819) HPK/GCK-LIKE KINASE O95819 51% 32 253 HGK. 74% 253 645 27% 6 149 92% 781 2019 HSAVA08 580870 388 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 57% 949 896 PROTEIN. 42% 926 792 63% 796 764 66% 1059 934 HSAWZ40 634000 391 WUblastx.64 (O00549) ORF2-LIKE PROTEIN O00549 64% 951 610 (FRAGMENT). 60% 613 8 HSDZM54 637870 393 WUblastx.64 NADH dehydrogenase (ubiquinone) pir|A00422|DNHUN3 88% 226 360 (EC 1.6.5.3) chain 3 - human mitochondrion HSHBF76 715838 394 WUblastx.64 (AAH08335) Unknown (protein for AAH08335 86% 762 457 IMAGE: 3506202) (Fra 73% 882 748 100% 1267 836 HSJBY32 702020 396 WUblastx.64 (Q9GZZ6) NEURONAL NICOTINIC Q9GZZ6 81% 466 639 ACETYLCHOLINE ALPHA10 57% 215 514 SUBUNIT PRECURSOR ( HSLHX15 777861 399 WUblastx.64 catalase (EC 1.11.1.6) - pir|I40767|I40767 86% 162 76 Campylobacter jejuni HSNBM34 635131 402 WUblastx.64 acyl-CoA dehydrogenase (EC 1.3.99.—) pir|S54183|S54183 84% 1548 1979 very-long-chain specific - human 100% 251 1546 HSOAH16 827058 403 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 57% 682 623 CLONE KAIA0536. 81% 624 544 68% 524 384 HSQDO85 853393 405 WUblastx.64 (Q9VCK0) CG10161 PROTEIN. Q9VCK0 67% 485 988 60% 60 521 56% 10 57 HSQES57 831222 406 WUblastx.64 (Q96EW4) Unknown (protein for Q96EW4 94% 195 980 MGC: 19936). HSRBE06 871264 407 WUblastx.64 (Q9H387) PRO2550. Q9H387 70% 1608 1327 HSSDI26 560722 408 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 68% 1398 1264 FOR MGC: 5149). HSSEA64 853395 409 WUblastx.64 (Q9HBT2) HYPOTHETICAL 17.2 KDA Q9HBT2 98% 7 243 PROTEIN. HSSEF77 658725 410 WUblastx.64 (O95637) WW DOMAIN BINDING O95637 42% 10 246 PROTEIN-1. 83% 296 829 HSSFE38 742512 411 HMMER PFAM: Ribonuclease HII PF01351 76.3 184 −142 2.1.1 WUblastx.64 (O75792) RIBONUCLEASE HI RNHL_HUMAN 91% 156 635 LARGE SUBUNIT (EC 3.1.26.—) 99% 587 1051 (RNASE HSWBE76 751308 413 WUblastx.64 (Q9NW15) CDNA FLJ10375 FIS, Q9NW15 100% 126 266 CLONE NT2RM2001950. HSXCP38 895392 414 WUblastx.64 hydroxymethylglutaryl-CoA lyase (EC pir|B45470|B45470 70% 17 895 4.1.3.4) - chicken HSYBI06 740766 415 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 69% 916 954 PROTEIN. 78% 821 913 HT5GR59 801930 420 WUblastx.64 (O60496) DOCKING PROTEIN. O60496 72% 70 1284 HTAEI78 637684 421 WUblastx.64 (Q9UKQ2) ADAM 28 PRECURSOR AD28_HUMAN 90% 85 174 (EC 3.4.24.—) (A DISINTEGRIN AND HTDAA78 566861 422 WUblastx.64 (Q9D8E7) 5830443F10RIK Q9D8E7 58% 84 302 PROTEIN. HTEAG62 812332 423 WUblastx.64 (Q9Y5Z7) HOST CELL FACTOR 2. Q9Y5Z7 60% 1 57 93% 14 2011 30% 107 631 HTECB02 806305 424 WUblastx.64 (AAK39520) BTB domain protein AAK39520 95% 33 1211 (Fragment). HTECC15 866488 425 WUblastx.64 (Q92558) WISKOTT-ALDRICH WAS1_HUMAN 95% 321 1100 SYNDROME PROTEIN FAMILY 70% 1525 1998 MEMBER 1 ( 89% 1105 1281 HTEDS12 838621 428 WUblastx.64 (Q9H0K0) HYPOTHETICAL 81.8 KDA Q9H0K0 97% 1029 1391 PROTEIN. 42% 1269 1490 100% 16 1011 HTEEF26 789606 431 WUblastx.64 (Q9H7X7) CDNA FLJ14117 FIS, Q9H7X7 81% 80 634 CLONE MAMMA1001785. HTEEF26 879704 432 WUblastx.64 (Q9H7X7) CDNA FLJ14117 FIS, Q9H7X7 81% 80 634 CLONE MAMMA1001785. HTEEW69 764835 433 WUblastx.64 (Q9Z1H7) GSG1. Q9Z1H7 65% 850 927 85% 707 769 50% 519 662 66% 908 943 65% 182 544 HTEGS07 827700 434 WUblastx.64 (Q9D143) 1110030K22RIK Q9D143 96% 183 593 PROTEIN. HTEHA56 806461 436 WUblastx.64 (Q9H9A0) CDNA FLJ12895 FIS, Q9H9A0 94% 2 217 CLONE NT2RP2004187, WEAKLY 65% 70 468 SIMILAR TO ZIN HTEJD29 695798 438 WUblastx.64 (Q60713) REVERSE Q60713 42% 1115 1285 TRANSCRIPTASE. 47% 874 1089 HTEMQ17 840387 440 WUblastx.64 (Q9D4P8) 4930579G24RIK Q9D4P8 90% 120 359 PROTEIN. HTENR63 877952 441 WUblastx.64 (Q9HD71) HYPOTHETICAL Q9HD71 33% 1278 1358 NUCLEAR FACTOR SBBI22. 78% 26 1168 HTGGM44 842856 442 WUblastx.64 probable phosphodiesterase I (EC pir|T43461|T43461 100% 1400 1924 3.1.4.1) - human (fragment) 83% 1925 2488 HTLBT80 840045 445 WUblastx.64 (Q9NQQ7) BA394O2.1 (CGI-15 Q9NQQ7 76% 1214 1405 PROTEIN). 74% 804 1223 47% 780 845 78% 313 825 HTLDA84 686397 446 WUblastx.64 (Q9H387) PRO2550. Q9H387 79% 1265 1134 60% 1442 1398 65% 1398 1243 HTLDN29 790195 447 WUblastx.64 (Q9CWL8) 5730471K09RIK Q9CWL8 96% 15 1226 PROTEIN. HTLEC82 811992 449 WUblastx.64 (Q99MI0) CELL GROWTH Q99MI0 98% 111 455 REGULATOR FALKOR. HTLEM16 779133 450 WUblastx.64 (O95638) WW DOMAIN BINDING O95638 92% 50 541 PROTEIN-2. 28% 987 1142 48% 617 841 HTLEV48 723799 451 WUblastx.64 (BAB55550) Bk125H2.1 protein. BAB55550 94% 10 825 HTLFA13 535937 452 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 57% 1118 873 HTLGI89 835069 454 WUblastx.64 (Q9BXS5) CLATHRIN- Q9BXS5 98% 104 682 ASSOCIATED PROTEIN AP47. 99% 675 1370 HTLIF11 843506 455 WUblastx.64 (Q9I8S4) ORNITHINE Q9I8S4 68% 309 356 DECARBOXYLASE-2. 59% 353 1687 HTLIF12 834946 456 WUblastx.64 (Q9DAR1) 1700001K04RIK Q9DAR1 47% 291 752 PROTEIN. HTLIF12 842691 457 WUblastx.64 (Q9DAR1) 1700001K04RIK Q9DAR1 47% 293 754 PROTEIN. HTLIF12 870167 458 WUblastx.64 (Q9DAR1) 1700001K04RIK Q9DAR1 47% 293 754 PROTEIN. HTLIF12 886780 459 WUblastx.64 (Q9DAR1) 1700001K04RIK Q9DAR1 47% 293 754 PROTEIN. HTLIF12 891533 460 WUblastx.64 (Q9DAR1) 1700001K04RIK Q9DAR1 47% 293 754 PROTEIN. HTLIF12 901225 461 WUblastx.64 (Q9DAR1) 1700001K04RIK Q9DAR1 47% 293 754 PROTEIN. HTNBK13 831967 463 WUblastx.64 (Q9Y3M2) HYPOTHETICAL 14.5 KDA Q9Y3M2 81% 123 500 PROTEIN. HTOAM11 664508 465 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 77% 428 363 CLONE LNG09295. 75% 586 425 HTOEV16 853616 468 WUblastx.64 (Q9NRZ5) 1-ACYL-SN- PLCD_HUMAN 98% 201 383 GLYCEROL-3-PHOSPHATE 95% 379 1164 ACYLTRANSFERASE DEL HTOHO21 732808 470 WUblastx.64 P47 LBC oncogene - human pir|I38434|I38434 97% 581 438 HTOHQ05 853621 471 WUblastx.64 (Q9UII4) CYCLIN-E BINDING Q9UII4 100% 669 791 PROTEIN 1. HTOJL95 806212 472 WUblastx.64 (Q15605) ORF1 CODES FOR A 40 KDA Q15605 86% 192 61 PRODUCT. 57% 876 730 57% 751 161 HTOJL95 762851 473 WUblastx.64 (Q15401) LINE-1 REPEAT MRNA Q15401 36% 683 609 WITH 2 OPEN READING FRAMES. 59% 966 820 71% 607 248 HTPDU17 840596 474 WUblastx.64 (Q9NW00) CDNA FLJ10404 FIS, Q9NW00 80% 553 1308 CLONE NT2RM4000486. 64% 1143 1664 HTSFJ32 637720 475 WUblastx.64 (Q9WUW2) VESICLE Q9WUW2 64% 747 788 ASSOCIATED MEMBRANE 94% 448 609 PROTEIN 2B. HTTCB60 853401 476 WUblastx.64 (Q9HAW0) RNA POLYMERASE III Q9HAW0 90% 6 881 TRANSCRIPTION INITIATION FACTOR BRFU. HTTEE41 840950 477 WUblastx.64 (P78371) T-COMPLEX PROTEIN 1, TCPB_HUMAN 98% 92 1696 BETA SUBUNIT (TCP-1-BETA) (CC HTTEZ02 702027 478 WUblastx.64 (Q9UEZ7) MAKORIN 1. Q9UEZ7 56% 278 346 98% 6 272 HTWEH94 561680 479 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 60% 1150 929 PROTEIN. HTXDC38 801935 482 WUblastx.64 (Q9BTX3) SIMILAR TO HSPC171 Q9BTX3 99% 100 573 PROTEIN. HTXDC77 844258 483 HMMER PFAM: Class I Histocompatibility PF00129 103.3 137 259 2.1.1 antigen, domains alpha 1 and 2 WUblastx.64 (P03989) HLA CLASS I 1B14_HUMAN 63% 880 945 HISTOCOMPATIBILITY ANTIGEN, 71% 65 256 B-27 ALPHA 80% 282 863 HTXFA72 853410 487 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 59% 1688 1557 PRODUCT. 66% 1839 1681 HTXKF95 834438 489 WUblastx.64 (AAH08360) Similar to hypothetical AAH08360 85% 233 553 protein FLJ22376 100% 2 112 HTXMZ07 834881 490 WUblastx.64 (Q9BRF3) SIMILAR TO RIKEN Q9BRF3 90% 3 1469 CDNA 2810468K17 GENE. HUFCL31 801938 491 WUblastx.64 (Q9D311) 9030623N16RIK Q9D311 60% 280 1224 PROTEIN. HUKBT67 844446 492 WUblastx.64 (BAB55428) CDNA FLJ14975 fis, BAB55428 100% 1040 1216 clone THYRO1001405, w 100% 8 61 30% 80 241 HUKDY82 570896 494 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 59% 1405 1145 CLONE MAMMA1000472. HUSCJ14 894699 495 WUblastx.64 tex261 protein - mouse pir|S47481|S47481 99% 74 661 HUSGL67 792637 496 WUblastx.64 (Q9Y2G2) CARD DOMAIN CRD8_HUMAN 100% 347 421 PROTEIN 8 (APOPTOTIC PROTEIN 65% 947 1006 NDPP1) (D 97% 469 954 HUSGU40 684975 497 WUblastx.64 (Q9BX98) UBIQUITIN A-52 Q9BX98 75% 840 433 RESIDUE RIBOSOMAL PROTEIN FUSION PRODUCT 1 (F HUVDJ48 564853 499 WUblastx.64 SHORT ISOFORM OF Q9P2N4 sp_vs|Q9P2N4- 92% 1510 1668 01|Q9P2N4 HWAAI12 830432 500 WUblastx.64 (Q9BWW4) SINGLE STRANDED Q9BWW4 82% 512 829 DNA BINDING PROTEIN-1. 87% 92 394 69% 941 1252 36% 521 685 37% 752 826 HWBCN36 722259 502 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 69% 1007 900 PROTEIN. 57% 887 846 HWBDJ08 762860 503 WUblastx.64 probable pol polyprotein-related pir|S21348|S21348 47% 901 833 protein 4 - rat 43% 1262 1131 53% 1134 904 HWDAC26 821335 505 WUblastx.64 (Q14287) HYPOTHETICAL Q14287 51% 1316 1471 PROTEIN (FRAGMENT). 57% 1093 1323 HWDAG96 796743 506 WUblastx.64 (AAH01119) Integrin beta 4 binding AAH01119 100% 108 842 protein. HWHPB78 740778 508 WUblastx.64 (Q9BUK4) SIMILAR TO Q9BUK4 61% 360 614 HYPOTHETICAL PROTEIN 100% 677 817 FLJ10709. HYABC84 789854 509 WUblastx.64 (Q99L03) SIMILAR TO TRP4- Q99L03 89% 209 553 ASSOCIATED PROTEIN TAP1 (FRAGMENT). HYABC84 865064 510 WUblastx.64 (Q9H429) DJ756N5.2 (A NOVEL Q9H429 92% 163 618 PROTEIN (DKFZP727M231) SIMILAR TO TRP4-AS H2CBD20 570796 511 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 42% 758 988 PROTEIN (FRAGMENT). 51% 987 1223 H2CBH91 826669 512 WUblastx.64 (Q9NXA3) CDNA FLJ20357 FIS, Q9NXA3 63% 133 222 CLONE HEP16545. 53% 3 125 75% 603 737 68% 263 643 H2LBA54 684290 513 WUblastx.64 (O60875) APOPTOSIS SPECIFIC O60875 97% 172 996 PROTEIN (DJ134E15.2) (APOPTOSIS SPECIFIC H2LBB09 658667 514 WUblastx.64 (Q9P0R6) HSPC210. Q9P0R6 95% 465 536 97% 73 477 H2LBB09 830636 515 WUblastx.64 (Q9P0R6) HSPC210. Q9P0R6 100% 504 575 97% 112 516 H2MBF60 695714 518 WUblastx.64 transcription factor TFIID 32K chain pir|I39141|I39141 100% 163 909 TAFII32 - human H6EEA48 847111 520 WUblastx.64 (AAH07644) Similar to RIKEN cDNA AAH07644 90% 478 933 9430029K10 gene (F 96% 27 482 H6EEN71 829201 521 WUblastx.64 (Q9BW90) SIMILAR TO Q9BW90 100% 1943 1779 PEROXISOME BIOGENESIS FACTOR 10. H6EEO05 865424 522 HMMER PFAM: EF hand PF00036 25 328 414 2.1.1 WUblastx.64 hypothetical protein pir|T17225|T17225 97% 265 522 DKFZp564C246.1 - human HACBJ11 797625 528 WUblastx.64 band-6-protein - human pir|S60712|S60712 100% 8 139 HACBS86 603946 529 WUblastx.64 (Q9BQB1) HYPOTHETICAL 22.5 KDA Q9BQB1 66% 89 682 PROTEIN. HADCL19 599065 535 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 57% 1070 780 CLONE COL04765. HADDC04 601695 537 WUblastx.64 (Q9GML5) HYPOTHETICAL 8.0 KDA Q9GML5 57% 2522 2379 PROTEIN. HADDP51 853356 539 HMMER PFAM: TBC domain PF00566 52.3 357 509 2.1.1 WUblastx.64 (Q9H695) CDNA: FLJ22474 FIS, Q9H695 99% 12 701 CLONE HRC10568. HADET62 607615 541 WUblastx.64 retrovirus-related hypothetical protein pir|S23650|S23650 33% 506 330 II - human 1 27% 325 77 50% 657 592 44% 808 668 HADEY08 799507 542 WUblastx.64 (Q9NX94) CDNA FLJ20367 FIS, Q9NX94 98% 663 1076 CLONE HEP18101. HADEY22 861628 544 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 77% 590 459 CLONE COL04765. HADFB84 668229 545 WUblastx.64 (Q9H387) PRO2550. Q9H387 67% 758 1009 HADFD10 843934 547 WUblastx.64 (Q9P1C6) PRO2738. Q9P1C6 52% 1254 1054 HADFW20 599066 550 WUblastx.64 (Q9H9H0) CDNA FLJ12759 FIS, Q9H9H0 83% 1065 823 CLONE NT2RP2001347. HADFX10 741054 551 WUblastx.64 (O60448) NEURONAL THREAD O60448 45% 1225 1040 PROTEIN AD7C-NTP. 62% 726 646 35% 1221 808 39% 1194 850 56% 917 729 33% 965 729 57% 1022 939 45% 726 628 51% 972 772 59% 972 715 57% 1225 1169 34% 1188 940 HADFY80 654831 552 WUblastx.64 (Q9H8X9) CDNA FLJ13153 FIS, Q9H8X9 94% 2 169 CLONE NT2RP3003409, WEAKLY SIMILAR TO HUM HADXA10 772423 555 WUblastx.64 (Q9NZ51) NEUROENDOCRINE Q9NZ51 78% 132 311 DIFFERENTIATION FACTOR. 79% 295 798 HADXA10 859777 556 WUblastx.64 (Q9D792) 9130011K15RIK Q9D792 49% 173 472 PROTEIN. HAFBB15 608180 557 HMMER PFAM: RNA 3′-terminal phosphate PF01137 159.5 186 440 2.1.1 cyclase WUblastx.64 RNA-3′-phosphate cyclase (EC pir|T48844|T48844 93% 6 446 6.5.1.4) 1 [validated] - human HAGAB62 588471 559 WUblastx.64 (Q9H2I4) DC42. Q9H2I4 77% 8 232 HAGAB83 823044 560 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 47% 1597 1472 CLONE KAT08285. 76% 1817 1599 HAGAF75 561933 562 WUblastx.64 (Q9UI59) PRO0478 PROTEIN. Q9UI59 77% 1203 1310 HAGAK40 731929 563 WUblastx.64 (Q9H387) PRO2550. Q9H387 80% 506 658 70% 707 838 HAGAZ36 564230 565 WUblastx.64 pro-pol-dUTPase polyprotein - murine pir|T29097|T29097 66% 613 578 endogenous retrovirus ERV-L 38% 897 655 (fragment) 34% 572 27 HAGBL31 679582 567 WUblastx.64 (Q13416) ORIGIN RECOGNITION ORC2_HUMAN 97% 509 610 COMPLEX SUBUNIT 2. HAGBO09 853357 568 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 78% 1533 1249 CLONE KAT08285. HAGBO12 601431 569 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 63% 348 241 PROTEIN. 76% 532 353 HAGBS89 846292 571 WUblastx.64 (Q9P195) PRO1722. Q9P195 56% 1125 841 HAGBV06 701966 572 WUblastx.64 (Q9UHC7) MAKORIN 1. Q9UHC7 66% 28 195 HAGBV25 838174 573 WUblastx.64 (Q9H7C8) CDNA: FLJ21040 FIS, Q9H7C8 99% 1 1644 CLONE CAE10642. HAGBV29 837203 574 WUblastx.64 (Q9H387) PRO2550. Q9H387 70% 2054 1761 71% 439 585 60% 309 455 56% 749 955 48% 732 836 73% 687 731 HAGCC87 638587 575 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 65% 992 1105 PROTEIN. 36% 54 116 57% 801 980 HAGCI69 560596 577 WUblastx.64 (Q9UI59) PRO0478 PROTEIN. Q9UI59 51% 1015 1185 HAGCZ70 747697 579 WUblastx.64 (Q9H397) PRO2852. Q9H397 41% 1672 1544 75% 1490 1359 77% 1366 1235 HAGDC73 724860 580 WUblastx.64 (Q9V662) CG12367 PROTEIN. Q9V662 37% 536 1006 HAGDJ53 821315 584 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 70% 1873 1781 PROTEIN. 46% 1796 1713 41% 1782 1582 HAGDL51 637488 586 WUblastx.64 (O60448) NEURONAL THREAD O60448 61% 1639 1532 PROTEIN AD7C-NTP. 70% 110 3 HAGDO70 812393 587 WUblastx.64 platelet-activating factor pir|JC4246|JC4246 94% 185 877 acetylhydrolase (EC 3.1.1.—) gamma chain - human HAGDT30 589514 588 WUblastx.64 (O75592) PROTEIN ASSOCIATED O75592 100% 11 280 WITH MYC. 24% 11 208 100% 1125 1946 25% 295 933 90% 511 1131 45% 1445 1504 HAGDW68 835631 589 WUblastx.64 (AAC14670) AAC14670 100% 390 470 WUGSC: H_DJ0651K02.1 protein 99% 2 385 (Fragment). HAGEK86 748222 592 WUblastx.64 (O43237) DYNEIN LIGHT DYJ2_HUMAN 88% 1066 1617 INTERMEDIATE CHAIN 2, CYTOSOLIC (LIC5 HAGEP30 604478 593 WUblastx.64 (Q9H387) PRO2550. Q9H387 84% 594 517 67% 788 606 HAGEQ67 838445 595 WUblastx.64 (Q94532) PUTATIVE TYPE III Q94532 55% 22 123 ALCOHOL DEHYDROGENASE. 59% 108 722 HAGEU26 608183 596 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 59% 356 481 PROTEIN. HAGFM58 604536 603 WUblastx.64 (Q9H3C0) PRO0898. Q9H3C0 62% 10 144 HAGFT48 780112 604 WUblastx.64 (Q9Y2Y7) FOOCEN-M (NOGO-B Q9Y2Y7 80% 761 1090 PROTEIN) (RTN-XS) (RETICULON 100% 1156 1281 4B). 48% 233 559 HAGFU31 751713 605 WUblastx.64 (O60448) NEURONAL THREAD O60448 65% 1214 1083 PROTEIN AD7C-NTP. 68% 1245 1084 45% 1093 1028 52% 1015 953 68% 1094 1029 71% 1230 967 HAGFW13 634611 606 WUblastx.64 (O00549) ORF2-LIKE PROTEIN O00549 65% 398 321 (FRAGMENT). 65% 453 385 70% 307 164 HAGHE85 838059 607 WUblastx.64 (Q9P195) PRO1722. Q9P195 58% 1665 1498 69% 1511 1365 HAHSD51 847014 612 WUblastx.64 (Q9D8B4) 2010012C24RIK Q9D8B4 50% 99 176 PROTEIN. 61% 182 505 HAIBV91 852223 615 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 59% 672 899 PRODUCT. 40% 1911 1955 HAICE62 834523 616 WUblastx.64 (Q9H3T5) MOB1 PROTEIN Q9H3T5 100% 203 850 (HYPOTHETICAL 25.1 KDA PROTEIN). HAICL90 637491 617 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 63% 716 438 PRODUCT. HAIDP45 847015 619 WUblastx.64 (AAH08590) Hypothetical 47.9 kDa AAH08590 100% 986 1096 protein. HAJAB88 780114 620 HMMER PFAM: Ribosomal protein L34 PF00468 32.3 276 401 2.1.1 WUblastx.64 (Q9BQ48) MITOCHONDRIAL Q9BQ48 98% 126 374 RIBOSOMAL PROTEIN L34 (L34MT) (UNKNOWN) (PROTE HAMGG01 783864 624 WUblastx.64 (Q9BY78) RING FINGER PROTEIN Q9BY78 86% 7 342 WITH LEUCINE ZIPPER RNF26. HANKC93 847018 626 WUblastx.64 (Q9H387) PRO2550. Q9H387 67% 648 565 52% 876 820 61% 748 656 HAPAD35 840584 627 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 76% 1346 1245 CLONE KAIA0536. 63% 1237 1064 HAPBR13 609976 628 WUblastx.64 (Q9H6T0) CDNA: FLJ21918 FIS, Q9H6T0 26% 273 452 CLONE HEP04006. 51% 95 274 40% 424 762 HAPBU09 762803 629 WUblastx.64 (O60859) NEUROPATHY TARGET O60859 100% 3 545 ESTERASE. HAPNJ33 835554 632 WUblastx.64 (BAA85159) Sec61. BAA85159 100% 1200 1403 98% 163 1230 HAPNL62 790340 633 WUblastx.64 (BAB55063) CDNA FLJ14456 fis, BAB55063 93% 1 1977 clone HEMBB1001915, m HAPNO50 834384 634 WUblastx.64 hypothetical protein pir|T08701|T08701 98% 27 332 DKFZp564N123.1 - human (fragment) 88% 3 29 86% 323 883 HAPPW83 847020 636 WUblastx.64 (Q9H387) PRO2550. Q9H387 77% 626 757 84% 475 627 HAPRK55 735887 643 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 63% 1379 1068 CLONE KAT08285. HAPSH37 847021 644 WUblastx.64 (Q9H387) PRO2550. Q9H387 75% 63 28 84% 199 68 HAQBY85 832384 646 WUblastx.64 (Q9D4H9) 4932409F11RIK Q9D4H9 100% 17 358 PROTEIN. 55% 1663 1716 28% 1066 1212 34% 1459 1728 93% 361 1539 HAQBZ15 801966 647 WUblastx.64 (AAH07201) Unknown (protein for AAH07201 100% 6 86 IMAGE: 2961284) (Fra 30% 1717 1824 93% 175 711 37% 1019 1564 29% 1277 1510 35% 1711 1836 97% 1705 1956 35% 843 953 31% 205 357 27% 187 408 67% 641 1708 52% 1711 1836 31% 849 962 31% 1078 1296 26% 1241 1696 36% 1123 1266 30% 1292 1564 27% 1081 1146 37% 882 953 30% 205 498 29% 1684 1938 29% 196 357 27% 1054 1317 90% 83 181 HARAE26 560598 650 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 65% 1204 1076 CLONE COL03536. 75% 1068 925 HARAT69 769389 651 WUblastx.64 (Q9VZ55) CG1582 PROTEIN. Q9VZ55 48% 2 1360 HARAZ81 832380 652 WUblastx.64 (Q9BQ36) MITOCHONDRIAL Q9BQ36 97% 7 327 RIBOSOMAL PROTEIN BMRP64 (HYPOTHETICAL 15.1 KD HASAU26 845848 653 WUblastx.64 (Q9H964) CDNA FLJ12984 FIS, Q9H964 94% 3 107 CLONE NT2RP3000047, WEAKLY 95% 132 194 SIMILAR TO NPL 98% 820 1425 40% 2405 2479 98% 188 829 100% 98 133 HASAY07 834511 655 WUblastx.64 catalase (EC 1.11.1.6) - pir|I40767|I40767 96% 263 177 Campylobacter jejuni HATAE01 654834 656 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 71% 569 507 CLONE KAT08285. 95% 636 574 72% 790 647 HATAL05 847023 658 WUblastx.64 (Q9ER61) MDM2 BINDING Q9ER61 68% 10 876 PROTEIN. HATCF80 780460 661 WUblastx.64 (Q99LV8) UNKNOWN (PROTEIN Q99LV8 78% 9 479 FOR IMAGE: 3489486) (FRAGMENT). HATCI67 847024 662 WUblastx.64 (Q9UHU3) PRO1659. Q9UHU3 85% 214 528 HATDH23 603959 668 WUblastx.64 (O43439) MTG8-LIKE PROTEIN O43439 98% 1 348 (MTG8 RELATED PROTEIN) (EHT) (EHT PROTEIN) HATDO84 609850 670 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 58% 798 646 CLONE KAIA0536. 77% 919 812 HATDU01 847028 671 WUblastx.64 (Q9H387) PRO2550. Q9H387 81% 1257 1225 58% 1198 974 HAUCC84 830672 677 WUblastx.64 (O75353) ANTI-DEATH PROTEIN. O75353 72% 44 259 98% 250 498 HAWAS41 877621 678 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 66% 754 930 PROTEIN. HAWBA65 542056 679 WUblastx.64 (Q24333) ELASTIN LIKE PROTEIN Q24333 100% 41 112 (FRAGMENT). HBAGH64 801884 680 WUblastx.64 (Q14287) HYPOTHETICAL Q14287 37% 810 568 PROTEIN (FRAGMENT). HBBBA42 841010 684 WUblastx.64 (Q9UIL1) HRIHFB2072 PROTEIN Q9UIL1 58% 3 263 (FRAGMENT). 57% 6 269 HBCAQ48 525002 690 WUblastx.64 (O75528) ADA3-LIKE PROTEIN. O75528 84% 73 345 HBGBE75 897455 694 WUblastx.64 (O60830) MITOCHONDRIAL I17B_HUMAN 97% 98 307 IMPORT INNER MEMBRANE TRANSLOCASE SU HBHAA53 603183 699 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 65% 439 326 PRODUCT. 68% 594 415 HBIAU43 840354 700 WUblastx.64 (Q9UI59) PRO0478 PROTEIN. Q9UI59 76% 1109 1225 HBIAW58 596805 701 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 66% 1268 1188 CLONE COL04765. 77% 1335 1270 68% 1486 1337 HBIBF26 845743 703 WUblastx.64 (Q9NQQ7) BA394O2.1 (CGI-15 Q9NQQ7 76% 1214 1405 PROTEIN). 74% 804 1223 47% 780 845 78% 313 825 HBIBQ69 580807 706 WUblastx.64 (BAB55217) CDNA FLJ14686 fis, BAB55217 94% 671 724 clone NT2RP2004961, m 100% 721 810 HBIBR38 612783 707 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 73% 802 503 CLONE KAT08285. HBIBS33 590280 709 WUblastx.64 (Q9H960) CDNA FLJ12988 FIS, Q9H960 62% 595 452 CLONE NT2RP3000080. HBIBZ20 688861 711 WUblastx.64 (O75717) ACIDIC O75717 99% 229 603 NUCLEOPLASMIC DNA-BINDING PROTEIN 1 (AND-1). HBICB80 637516 712 WUblastx.64 (Q9BV17) SIMILAR TO CG9172 Q9BV17 90% 545 637 GENE PRODUCT. 91% 18 557 HBJAC40 841235 713 WUblastx.64 (Q9P112) CHROMOSOME 16 OPEN Q9P112 100% 8 73 READING FRAME 5. 36% 5 70 57% 11 52 53% 85 180 100% 192 632 HBJAV56 603529 714 WUblastx.64 (Q9H387) PRO2550. Q9H387 58% 642 478 67% 715 623 75% 780 745 HBJBR40 581104 717 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 54% 931 860 PROTEIN. 66% 861 745 HBJCH46 609859 718 WUblastx.64 conserved hypothetical protein pir|D83014|D83014 53% 1070 900 PA5065 [imported] - Pseudomonas 78% 900 1 aeruginosa (strain PAO1) HBJCS26 821682 720 WUblastx.64 (Q9H0D4) HYPOTHETICAL 54.1 KDA Q9H0D4 92% 227 1576 PROTEIN. HBJDR18 604907 723 WUblastx.64 (Q9H6G8) CDNA: FLJ22294 FIS, Q9H6G8 81% 940 875 CLONE HRC04426. 77% 1092 934 HBJDR83 600395 724 WUblastx.64 (Q14273) POL/ENV ORF. Q14273 71% 559 621 60% 813 902 68% 614 718 41% 1223 1291 62% 14 556 HBJEL21 866158 726 WUblastx.64 (Q9UQR1) ZINC FINGER PROTEIN Z148_HUMAN 97% 807 2063 148 (ZINC FINGER DNA BINDING P HBJFH84 836997 727 WUblastx.64 (Q9LM25) T10O22.22. Q9LM25 30% 118 921 HBJFJ26 873844 3102 WUblastx.64 (Q9BWK5) UNKNOWN (PROTEIN Q9BWK5 94% 657 758 FOR MGC: 5242). 100% 336 389 100% 427 657 HBJHO83 610259 736 HMMER PFAM: ENV polyprotein (coat PF00429 40.5 207 389 2.1.1 polyprotein) WUblastx.64 (Q85641) 3′ END OF THE GENOME Q85641 85% 2 61 OF MOLONEY MURINE 28% 177 365 LEUKEMIA VIRUS (CODES 46% 64 180 HBJIR14 793391 741 WUblastx.64 (Q9H832) CDNA FLJ13968 FIS, Q9H832 91% 134 871 CLONE Y79AA1001493, WEAKLY SIMILAR TO UBI HBJJA26 743181 742 WUblastx.64 (Q9BRM8) UNKNOWN (PROTEIN Q9BRM8 47% 647 423 FOR MGC: 13219). HBJND04 837242 745 HMMER PFAM: Bacterial mutT protein PF00293 26.2 −360 −479 2.1.1 WUblastx.64 (Q9BW91) UNKNOWN (PROTEIN Q9BW91 91% 309 1352 FOR MGC: 3037). HBJND57 612785 746 WUblastx.64 (Q9VCB4) CG17741 PROTEIN. Q9VCB4 52% 1 318 HBKEE60 793788 749 WUblastx.64 pro-pol-dUTPase polyprotein - murine pir|T29097|T29097 64% 97 204 endogenous retrovirus ERV-L 39% 3 128 (fragment) 62% 273 620 HBKEI41 827278 750 WUblastx.64 (BAB47402) CG10671-like. BAB47402 100% 436 465 90% 596 757 59% 498 623 77% 5 436 HBMBD51 842176 751 WUblastx.64 hypothetical protein pir|T50632|T50632 98% 2197 2709 DKFZp762E1511.1 - human (fragment) HBMBD73 839564 752 WUblastx.64 MAP kinase 1 (EC 2.7.1.—) - human pir|JQ1400|JQ1400 91% 93 161 92% 127 1137 HBMBM17 637518 754 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 75% 950 840 CLONE KAIA0536. 75% 1083 949 HBMCL59 608668 755 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 45% 630 472 CLONE COL04765. 60% 796 608 HBMCM96 821318 756 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 57% 2253 2155 CLONE COL04765. 63% 2177 1971 HBMCQ74 856461 757 WUblastx.64 (Q9JKP5) MUSCLEBLIND. Q9JKP5 100% 561 590 52% 13 213 83% 1 465 HBMCQ74 864382 758 WUblastx.64 (Q9JKP5) MUSCLEBLIND. Q9JKP5 100% 561 590 52% 13 213 83% 1 465 HBMDM08 837927 760 WUblastx.64 (Q9P0I2) 30 KDA PROTEIN. Q9P0I2 94% 190 972 HBMSO30 843389 762 WUblastx.64 (Q9D4B1) 4933405A16RIK Q9D4B1 78% 52 174 PROTEIN. HBMTM50 609988 763 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 76% 763 689 CLONE KAIA0536. 55% 962 765 HBMUD59 701970 764 WUblastx.64 (Q9BR03) C367G8.2 (NOVEL Q9BR03 91% 9 647 PROTEIN) (FRAGMENT). HBMUR39 647594 767 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 55% 883 803 PRODUCT. 39% 810 577 HBMWC39 523713 770 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 71% 1055 1117 CLONE COL04765. 67% 820 1053 HBMWS52 872553 772 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 62% 1341 997 CLONE COL04765. HBMXG01 689522 774 WUblastx.64 (Q9BGX7) HYPOTHETICAL 13.0 KDA Q9BGX7 58% 838 885 PROTEIN. 71% 900 1055 HBMXG76 580812 775 WUblastx.64 probable phosphodiesterase I (EC pir|T43461|T43461 98% 37 222 3.1.4.1) - human (fragment) HBMXW83 725335 777 WUblastx.64 glutamate- cysteine ligase (EC 6.3.2.2) pir|JH0611|JH0611 100% 492 584 heavy chain - human 90% 373 498 HBNAE74 637524 778 WUblastx.64 (Q9NU78) DJ622L5.7.1 (NOVEL Q9NU78 100% 6 146 PROTEIN (ISOFORM 1)). HBNAX16 843727 779 WUblastx.64 (Q9BT21) SIMILAR TO RIKEN Q9BT21 92% 360 2639 CDNA 2610005L19 GENE (FRAGMENT). HBODK40 852382 781 WUblastx.64 (Q9Y6B7) ADAPTER-RELATED A4B1_HUMAN 89% 55 696 PROTEIN COMPLEX 4 BETA 1 99% 674 1741 SUBUNIT ( HBODV76 866420 782 WUblastx.64 (O60662) KELCH-RELATED KRP1_HUMAN 83% 474 902 PROTEIN 1 (KEL-LIKE PROTEIN 100% 71 475 23) (SAR 96% 808 1905 HBPAF39 850786 784 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 51% 1217 849 CLONE KAIA0536. HBQAC72 799512 786 WUblastx.64 transcription factor znf6 - human pir|S25409|S25409 90% 2072 1110 33% 1844 1227 30% 2024 1215 30% 1757 1194 31% 1844 1401 HBSAJ63 848683 788 WUblastx.64 (BAB55144) CDNA FLJ14576 fis, BAB55144 92% 11 406 clone NT2RM4001092, w HBSDD24 839802 790 WUblastx.64 (Q9UJ70) N- Q9UJ70 100% 886 1251 ACETYLGLUCOSAMINE KINASE PROTEIN (EC 2.7.1.59). HBWBD25 800765 791 WUblastx.64 (O08872) PUTATIVE RNA O08872 49% 850 698 BINDING PROTEIN 1 30% 1473 853 (FRAGMENT). HBXAS93 836513 792 WUblastx.64 (Q14940) SODIUM/HYDROGEN NAH5_HUMAN 85% 532 1023 EXCHANGER 5 (NA(+)/H(+) 78% 2 238 EXCHANGER 90% 256 321 HBXAW57 815650 794 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 56% 1223 1071 PRODUCT. HBXBM78 812527 797 WUblastx.64 (Q9Y613) FH1/FH2 DOMAINS- FHOS_HUMAN 84% 846 259 CONTAINING PROTEIN (FORMIN 28% 2503 2138 HOMOLOG 100% 1004 852 97% 1903 1046 100% 2311 2042 42% 2184 2122 28% 1777 1334 55% 2520 2248 HBXCD59 860439 798 WUblastx.64 (Q9C026) TRIPARTITE MOTIF Q9C026 99% 59 823 PROTEIN TRIM9 ISOFORM BETA. HBXCG08 628501 800 WUblastx.64 (Q9BSG3) UNKNOWN (PROTEIN Q9BSG3 98% 758 1024 FOR MGC: 12974). HBXCM52 799513 801 WUblastx.64 (Q9BH03) HYPOTHETICAL 12.0 KDA Q9BH03 53% 683 850 PROTEIN. 73% 784 942 80% 903 1028 HBXCQ03 589516 802 WUblastx.64 (Q9Y296) PTD009 (HSPC172). Q9Y296 100% 382 672 100% 17 382 HBXDN08 566765 806 WUblastx.64 (O43597) SPROUTY HOMOLOG 2 SPY2_HUMAN 100% 8 256 (SPRY-2). HBXDN65 840021 807 WUblastx.64 (Q9H387) PRO2550. Q9H387 75% 1925 1641 HBXFA04 842901 808 WUblastx.64 (AAH08467) Similar to RIKEN cDNA AAH08467 95% 287 499 1110001J03 gene. HBXFE64 838824 809 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 57% 1646 1320 CLONE COL04765. HBXFP72 688040 811 WUblastx.64 (Q9H754) CDNA: FLJ21308 FIS, Q9H754 93% 7 1155 CLONE COL02131. HBXFS31 815651 812 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 65% 469 678 CLONE KAT08285. 70% 1551 1261 HBXFW01 847001 813 HMMER PFAM: Stathmin family PF00836 185.7 266 523 2.1.1 WUblastx.64 (Q9H169) RB3 PROTEIN Q9H169 100% 523 690 (HYPOTHETICAL 22.1 KDA 100% 125 523 PROTEIN). HBXGE12 1310891 814 WUblastx.64 (O95902) UNKNOWN O95902 100% 31 411 (FRAGMENT). HBXGE12 745398 3103 WUblastx.64 (AF131851) Unknown [Homo sapiens] gb|AAD20061.1| 100% 10 390 HBXGL91 901845 815 WUblastx.64 (Q9H2J7) ORPHAN Q9H2J7 100% 949 1470 NEUROTRANSMITTER 100% 10 99 TRANSPORTER V7-3. 95% 389 979 HBXGM24 821320 816 WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 77% 712 647 human 46% 740 660 48% 639 475 57% 85 23 64% 530 84 HCBAB34 847002 821 WUblastx.64 (Q9BWF8) UNKNOWN (PROTEIN Q9BWF8 95% 1009 1308 FOR IMAGE: 3355813) 98% 814 963 (FRAGMENT). 95% 680 823 60% 29 583 HCDAH02 653066 825 WUblastx.64 (BAB55208) CDNA FLJ14668 fis, BAB55208 81% 655 491 clone NT2RP2003194. 83% 492 241 HCDAP33 566794 826 WUblastx.64 (O75964) ATP SYNTHASE G ATPN_HUMAN 73% 407 664 CHAIN, MITOCHONDRIAL (EC 3.6.1.34) HCDAR40 654821 827 WUblastx.64 (Q9P195) PRO1722. Q9P195 63% 837 514 HCDAS02 896667 828 WUblastx.64 (O75153) PUTATIVE EUKARYOTIC IF3X_HUMAN 89% 79 867 TRANSLATION INITIATION FACTOR HCDBO32 831942 830 WUblastx.64 (AAH17472) Hypothetical 21.3 kDa AAH17472 69% 643 801 protein. 100% 239 583 HCDBW67 733860 831 WUblastx.64 (Q9H410) DJ469A13.2 (NOVEL Q9H410 97% 43 255 PROTEIN) (FRAGMENT). HCDCB03 571037 833 HMMER PFAM: Zinc carboxypeptidase PF00246 103.3 35 337 2.1.1 WUblastx.64 carboxypeptidase H (EC 3.4.17.10) pir|S09489|S09489 91% 657 728 precursor - human 66% 11 664 HCDCE51 813504 834 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 55% 1016 837 PRODUCT. HCDCI42 847004 835 WUblastx.64 (Q9UHD2) TANK BINDING Q9UHD2 100% 678 875 KINASE TBK1 (NF-KB- ACTIVATING KINASE NAK). HCDDB15 841041 836 WUblastx.64 (Q9BW53) UNKNOWN (PROTEIN Q9BW53 100% 98 703 FOR IMAGE: 3343149) (FRAGMENT). HCDDY28 892137 838 WUblastx.64 (Q14287) HYPOTHETICAL Q14287 55% 1458 1312 PROTEIN (FRAGMENT). 60% 1312 1013 HCDEB19 587264 839 WUblastx.64 nuclear matrix protein NMP 238 - pir|JE0334|JE0334 100% 1218 1436 human 93% 426 470 99% 473 829 98% 76 438 31% 605 700 31% 1335 1430 91% 838 1218 HCDES69 609997 841 WUblastx.64 (Q9H8N2) CDNA FLJ13381 FIS, Q9H8N2 61% 524 423 CLONE PLACE1001010. 62% 687 553 HCE1D45 664481 842 WUblastx.64 (Q9UDU0) WUGSC: H_DJ400N23.1 Q9UDU0 84% 10 279 PROTEIN (ZINC FINGER SARCOMA GENE LONG A HCE1T53 843680 844 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 71% 725 663 CLONE KAT08285. 65% 843 739 92% 1450 1412 67% 1409 1140 HCE1Y27 637529 845 WUblastx.64 (O95741) COPINE VI (NEURONAL- CNE6_HUMAN 80% 887 946 COPINE) (N-COPINE). 98% 519 896 95% 901 1353 93% 16 561 HCE1Y34 809084 846 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 61% 1718 1680 CLONE KAIA0536. 51% 1680 1411 HCE2E47 886155 848 WUblastx.64 (Q9H387) PRO2550. Q9H387 72% 1254 1448 67% 1450 1533 HCE2P90 737935 850 WUblastx.64 (AAH07558) Unknown (protein for AAH07558 47% 792 896 MGC: 15483). 42% 640 795 HCE3C46 737889 852 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 41% 186 260 CLONE KAT08285. 62% 751 1032 HCE3D58 873227 853 WUblastx.64 (Q9H026) HYPOTHETICAL 16.0 KDA Q9H026 70% 421 879 PROTEIN (FRAGMENT). HCE3N23 810211 857 WUblastx.64 (Q9D2L0) 4833417L20RIK Q9D2L0 42% 1150 1539 PROTEIN. 38% 1147 1536 71% 423 590 70% 913 1044 55% 584 925 54% 916 1047 44% 370 423 58% 863 913 72% 77 358 HCE3R01 834836 858 WUblastx.64 (O95251) HISTONE O95251 76% 6 320 ACETYLTRANSFERASE. 84% 68 1111 HCE3R01 841472 859 WUblastx.64 (O95251) HISTONE O95251 76% 6 320 ACETYLTRANSFERASE. 84% 68 1111 HCE3R01 844574 860 WUblastx.64 (O95251) HISTONE O95251 76% 6 320 ACETYLTRANSFERASE. 84% 68 1111 HCE3R46 844450 861 WUblastx.64 (O95894) UNKNOWN (DERP2). O95894 91% 131 1165 HCE4H32 843941 862 WUblastx.64 (Q9NVF5) CDNA FLJ10769 FIS, Q9NVF5 92% 926 964 CLONE NT2RP4000151. 65% 964 1104 64% 73 402 94% 333 893 HCE4H32 874256 863 WUblastx.64 (Q9P0P1) HSPC237. Q9P0P1 100% 1260 1322 100% 1325 1765 HCE4W88 792953 865 WUblastx.64 (Q9Y5W4) MLL SEPTIN-LIKE Q9Y5W4 100% 3676 2003 FUSION PROTEIN (CELL DIVISION CONTROL PROTEI HCE5B62 566864 866 WUblastx.64 (O60448) NEURONAL THREAD O60448 70% 1011 850 PROTEIN AD7C-NTP. 47% 1056 757 64% 1653 1486 58% 1563 1441 47% 953 810 70% 1470 1420 34% 820 725 40% 1023 943 62% 956 855 58% 778 728 58% 1553 1425 63% 1653 1546 61% 1632 1579 38% 1631 1500 36% 981 748 55% 1696 1637 56% 1505 1410 50% 1496 1431 45% 823 725 59% 846 781 61% 995 834 43% 1557 1420 84% 1683 1645 62% 1673 1425 HCE5H86 847032 867 WUblastx.64 (O95637) WW DOMAIN BINDING O95637 83% 1574 2107 PROTEIN-1. 66% 847 1248 66% 1300 1539 HCE5J64 688883 868 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 64% 2021 2122 PROTEIN. 56% 1903 2031 HCEBF54 847033 869 WUblastx.64 (Q9NQ43) HYPOTHETICAL 18.4 KDA Q9NQ43 100% 338 583 PROTEIN (FRAGMENT). HCEDN07 847034 874 WUblastx.64 (Q9H387) PRO2550. Q9H387 93% 1256 1212 86% 1186 1121 71% 1410 1255 HCEEG48 896688 876 WUblastx.64 (AAK55521) PRO0764. AAK55521 70% 919 680 HCEEM33 821322 877 WUblastx.64 hypothetical protein pir|T46310|T46310 100% 3 746 DKFZp434G0511.1 - human HCEFA94 822850 882 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 56% 1066 1224 PROTEIN. HCEFG93 745400 884 WUblastx.64 (Q9H387) PRO2550. Q9H387 66% 1178 1134 80% 1114 1052 76% 1323 1186 HCEFH31 801890 885 WUblastx.64 (Q9ULW1) SPONDIN 2. Q9ULW1 100% 1503 1685 HCEFK56 872554 886 WUblastx.64 (Q9UIU2) DYNACTIN 1 P150 Q9UIU2 92% 7 2088 ISOFORM. HCEGK81 844452 890 WUblastx.64 (Q9H387) PRO2550. Q9H387 88% 524 649 68% 390 455 76% 487 525 HCEGS49 846298 891 WUblastx.64 (O60448) NEURONAL THREAD O60448 42% 711 529 PROTEIN AD7C-NTP. 92% 651 610 61% 856 710 72% 870 838 66% 721 659 61% 872 711 68% 857 579 HCEGY33 753258 893 WUblastx.64 (O75843) ADAPTER-RELATED A1G2_HUMAN 97% 1131 988 PROTEIN COMPLEX 1 GAMMA 2 89% 873 760 SUBUNIT 100% 1396 1274 68% 393 256 96% 603 523 98% 2024 1818 HCEHW24 560610 894 WUblastx.64 (Q9BGX7) HYPOTHETICAL 13.0 KDA Q9BGX7 55% 1001 1297 PROTEIN. HCEJL08 722208 895 WUblastx.64 (AAH08203) Chromosome X open AAH08203 59% 36 170 reading frame 12. 96% 581 775 58% 2 103 79% 130 570 HCELB04 847375 897 WUblastx.64 (Q9P290) POTENT BRAIN TYPE Q9P290 96% 994 1083 ORGANIC ION TRANSPORTER. 94% 156 263 71% 677 1024 100% 317 544 62% 1106 1177 73% 500 544 75% 1064 1579 HCEMA08 846468 898 WUblastx.64 ATPase inhibitor precursor, pir|JC7175|JC7175 65% 1599 1739 mitochondrial - human 100% 67 216 HCENQ22 740746 900 WUblastx.64 (Q9H926) CDNA FLJ13059 FIS, Q9H926 80% 1086 934 CLONE NT2RP3001589. HCEOF01 564906 901 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 50% 1669 1586 CLONE COL04765. 75% 1551 1366 HCEOF01 850521 902 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 50% 1668 1585 CLONE COL04765. 75% 1550 1365 HCEOV48 850681 905 WUblastx.64 (Q9DA75) 1700018O18RIK Q9DA75 83% 1344 1739 PROTEIN. 76% 193 1311 HCEPO08 637543 907 WUblastx.64 ribosomal protein L29, cytosolic - pir|S65784|S65784 100% 614 504 human 64% 516 145 HCESB03 812940 908 WUblastx.64 (Q9HD86) NAG18. Q9HD86 76% 729 857 HCETL19 702090 911 WUblastx.64 retrovirus-related env polyprotein pir|E24483|VCHUER 79% 580 464 pseudogene - human HCFAT42 815652 915 WUblastx.64 (Q9G6T1) CYTOCHROME Q9G6T1 79% 699 884 OXIDASE SUBUNIT 3. 47% 369 431 HCFAT66 821337 916 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 64% 1754 1662 CLONE HEMBA1006036. 69% 1641 1453 HCFBA30 608166 917 WUblastx.64 (Q9BU56) UNKNOWN (PROTEIN Q9BU56 75% 357 692 FOR IMAGE: 3940029) 45% 192 245 (FRAGMENT). HCFBM77 604576 918 WUblastx.64 (O60448) NEURONAL THREAD O60448 59% 729 664 PROTEIN AD7C-NTP. 75% 880 719 77% 865 605 HCFCB72 824165 920 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 63% 1537 1370 CLONE KAIA0536. 59% 1707 1555 HCFCG91 897509 921 WUblastx.64 (Q9V677) CG8858 PROTEIN. Q9V677 47% 53 511 32% 786 2780 43% 492 785 24% 480 950 23% 1269 1565 21% 1275 1643 26% 1800 1922 HCFCM81 847378 922 WUblastx.64 (O00398) PUTATIVE PURINERGIC O00398 96% 281 1297 RECEPTOR P2Y10. HCFLJ52 1307037 928 WUblastx.64 (O00466) K12 PROTEIN O00466 95% 362 418 PRECURSOR. 100% 415 561 100% 121 372 HCFLJ52 753260 3104 WUblastx.64 (AX055560) unnamed protein product emb|CAC22100.1| 100% 91 126 [Homo sapiens] 56% 14 94 98% 123 287 HCFLY20 786452 932 WUblastx.64 hypothetical protein pir|T46299|T46299 85% 57 380 DKFZp434J0310.1 - human 93% 437 784 65% 936 1022 40% 167 226 76% 757 936 HCFLY20 858875 933 WUblastx.64 hypothetical protein pir|T46299|T46299 85% 68 391 DKFZp434J0310.1 - human 72% 959 1033 40% 178 237 89% 448 951 HCFMX16 581042 938 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 54% 304 29 PROTEIN. HCFMX88 825989 939 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 71% 1068 1130 CLONE HEMBA1006036. 77% 853 945 69% 959 1066 HCFNM40 746864 940 WUblastx.64 catalase (EC 1.11.1.6) - pir|I40767|I40767 84% 246 130 Campylobacter jejuni HCFNM50 732010 941 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 75% 1258 1103 CLONE KAT08285. HCFNN75 762959 943 WUblastx.64 (Q9BGZ4) HYPOTHETICAL 11.6 KDA Q9BGZ4 56% 821 696 PROTEIN. HCGBA15 604603 946 WUblastx.64 (AAK55521) PRO0764. AAK55521 81% 791 726 80% 925 881 80% 724 662 65% 903 790 HCHAC68 610370 947 HMMER PFAM: Ank repeat PF00023 58.1 462 560 2.1.1 WUblastx.64 (Q9Y290) RELA ASSOCIATED Q9Y290 90% 201 770 INHIBITOR. 93% 52 96 100% 770 871 70% 6 50 HCHBP49 892141 948 WUblastx.64 hypothetical protein pir|T47147|T47147 91% 8 457 DKFZp761H229.1 - human (fragment) HCHCG33 862534 950 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 66% 2068 1988 CLONE COL04765. 75% 2285 2073 HCHMY57 833049 951 WUblastx.64 (Q9BV10) UNKNOWN (PROTEIN Q9BV10 84% 12 1202 FOR MGC: 3136). HCHOC06 688042 952 WUblastx.64 (Q9BZH7) GASTRIC CANCER- Q9BZH7 75% 131 424 RELATED PROTEIN VRG107. HCHOY52 837297 953 WUblastx.64 (Q9BT25) UNKNOWN (PROTEIN Q9BT25 89% 27 1223 FOR IMAGE: 3636299) (FRAGMENT). HCHQB93 793648 954 WUblastx.64 GTP-binding protein 2 - human pir|PC7084|PC7084 89% 767 985 (fragment) 92% 258 788 86% 2 46 37% 36 143 100% 61 258 HCHQB93 875853 955 WUblastx.64 GTP-binding protein 2 - human pir|PC7084|PC7084 89% 767 985 (fragment) 92% 258 788 86% 2 46 37% 36 143 100% 61 258 HCLCU75 862406 957 WUblastx.64 (Q24333) ELASTIN LIKE PROTEIN Q24333 95% 51 122 (FRAGMENT). HCMSA37 598712 958 WUblastx.64 (Q9UHS7) PRO1992. Q9UHS7 63% 902 795 HCMSR07 821338 959 WUblastx.64 (Q9UI59) PRO0478 PROTEIN. Q9UI59 91% 1388 1489 HCNSF01 901060 964 WUblastx.64 (Q9Y6N5) HYPOTHETICAL 50.0 KDA Q9Y6N5 100% 107 1456 PROTEIN. HCPAE41 799546 967 WUblastx.64 (Q9P0P0) HSPC238 Q9P0P0 81% 490 443 (HYPOTHETICAL 17.9 KDA 73% 444 31 PROTEIN). HCQAS72 828082 970 WUblastx.64 (AAK58423) PC2-glutamine-rich- AAK58423 91% 22 57 associated protein. 93% 1010 1054 100% 57 119 71% 122 463 HCQBM95 841011 971 WUblastx.64 ribosomal protein L39, cytosolic pir|JC4229|R6RT39 84% 409 447 [validated] - rat 88% 282 389 HCRAI29 844084 979 WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 34% 329 132 human 56% 48 1 39% 493 305 61% 89 12 HCRBL20 709660 981 WUblastx.64 (Q9NWM9) CDNA FLJ20730 FIS, Q9NWM9 94% 1 678 CLONE HEP10359. HCRBX84 1007105 982 WUblastx.64 (Q9H1F6) DJ453C12.6.1 Q9H1F6 91% 37 177 (UNCHARACTERIZED 63% 51 617 HYPOTHALAMUS PROTEIN (ISOFORM HCRMA24 897005 983 WUblastx.64 hypothetical protein pir|T08683|T08683 100% 388 591 DKFZp564J2123.1 - human (fragment) HCRMR35 849071 984 WUblastx.64 (Q9H175) HYPOTHETICAL 59.6 KDA Q9H175 48% 43 858 PROTEIN. HCRMR35 874743 985 WUblastx.64 (Q9H175) HYPOTHETICAL 59.6 KDA Q9H175 48% 43 858 PROTEIN. HCRMR35 897434 986 WUblastx.64 (Q9H175) HYPOTHETICAL 59.6 KDA Q9H175 49% 43 858 PROTEIN. HCROC18 884144 987 WUblastx.64 (Q9H8A5) CDNA FLJ13824 FIS, Q9H8A5 95% 7 1695 CLONE THYRO1000505. HCUAE53 665716 988 WUblastx.64 hypothetical protein [imported] - pir|E86188|E86188 58% 504 304 Arabidopsis thaliana 80% 225 76 56% 895 653 28% 542 459 62% 1299 1156 HCUAT74 834611 990 WUblastx.64 hypothetical protein UL126 - human pir|S09875|S09875 68% 3 191 cytomegalovirus (strain AD169) HCUBN69 826610 994 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 78% 831 763 FOR MGC: 5149). 72% 626 573 80% 760 626 HCUBY47 842787 995 WUblastx.64 (Q9EPL2) CALSYNTENIN-1 Q9EPL2 97% 444 313 PROTEIN PRECURSOR. 97% 1003 869 HCUCV66 806577 996 WUblastx.64 (BAB22833) 18 days embryo cDNA, BAB22833 100% 36 362 RIKEN full-length e HCUFC77 745374 1001 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 77% 1526 1461 CLONE KAIA0536. 61% 1449 1234 HCUFD17 847039 1002 WUblastx.64 (Q9HBZ6) HT005 PROTEIN. Q9HBZ6 100% 10 195 HCUFD46 757481 1003 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 43% 580 386 CLONE COL04765. HCUFX08 612843 1008 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 55% 1214 1128 PROTEIN. 65% 1329 1216 HCUHE27 562766 1014 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 61% 764 639 PRODUCT. HCWAK88 839083 1017 WUblastx.64 probable excinuclease ABC chain A - pir|T33732|T33732 86% 435 370 Zymomonas mobilis 31% 815 321 32% 1969 1814 64% 956 501 67% 362 42 57% 1237 1181 69% 1306 998 85% 1993 1538 HCWAL10 639023 1018 WUblastx.64 (Q9Y2N3) NUCLEAR ENVELOPE N121_HUMAN 100% 754 680 PORE MEMBRANE PROTEIN POM 100% 240 118 121 (PO HCWDM69 684527 1022 WUblastx.64 (Q9P195) PRO1722. Q9P195 66% 819 748 78% 886 818 70% 1001 900 HCWEB72 639041 1024 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 56% 872 798 CLONE KAIA0536. 48% 588 508 65% 757 617 HCWEI82 799516 1026 WUblastx.64 (Q9P1N7) PRO0974. Q9P1N7 47% 384 656 HCWEM96 812735 1027 WUblastx.64 probable ATP-binding component of pir|G83165|G83165 74% 1901 1146 ABC transporter PA3838 [imported] - Pseudomonas aeruginosa (strain PAO1) HCWHD30 834724 1031 WUblastx.64 (AAH07609) Similar to hypothetical AAH07609 86% 634 590 protein PRO1722. 80% 782 633 HCWHT34 692400 1032 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 68% 556 777 HCWHT52 834618 1033 WUblastx.64 hypothetical protein UL126 - human pir|S09875|S09875 75% 20 274 cytomegalovirus (strain AD169) HCWKO32 610619 1034 WUblastx.64 (Q9GMU5) HYPOTHETICAL 14.1 KDA Q9GMU5 82% 496 600 PROTEIN. HCWUW24 799517 1037 WUblastx.64 (Q9H387) PRO2550. Q9H387 45% 541 696 33% 60 245 77% 686 817 HCYBA32 847042 1038 WUblastx.64 (Q9H3F8) MSTP016. Q9H3F8 100% 329 718 79% 925 1800 HDABR74 861789 1040 WUblastx.64 (O60717) 7- O60717 81% 5 37 DEHYDROCHOLESTEROL 95% 69 908 REDUCTASE (EC 1.3.1.21) (FRAGMENT). HDFIB37 821339 1045 WUblastx.64 (Q9D8F2) 2010004A03RIK Q9D8F2 74% 792 884 PROTEIN. 64% 357 431 65% 388 816 HDHEA33 847044 1048 WUblastx.64 (Q9BU66) UNKNOWN (PROTEIN Q9BU66 40% 482 619 FOR MGC: 10433). 45% 159 638 82% 1054 1494 HDHEB12 610260 1049 WUblastx.64 (Q9NZ96) NEUROLIGIN 3 Q9NZ96 94% 2 697 ISOFORM HNL3S (FRAGMENT). HDLAL94 843583 1054 HMMER PFAM: ADP-ribosylation factor PF00025 309.2 195 728 2.1.1 family WUblastx.64 (Q9D4P0) 4930587A11RIK Q9D4P0 100% 192 728 PROTEIN. HDPAB86 901851 1055 WUblastx.64 (Q9NYH9) HEPATOCELLULAR Q9NYH9 56% 74 538 CARCINOMA-ASSOCIATED 97% 310 1863 ANTIGEN 66. HDPAE80 778068 1056 WUblastx.64 (Q9BR63) PHENYLALANYL-TRNA Q9BR63 93% 1435 1662 SYNTHETASE BETA-SUBUNIT 78% 1604 1744 (FRAGMENT). 54% 383 661 32% 417 761 98% 905 1438 HDPAQ86 612855 1057 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 48% 1419 1246 CLONE COL03536. 76% 1250 1122 HDPBN48 838593 1059 WUblastx.64 (Q9H013) MELTRIN-BETA/ADAM Q9H013 84% 525 773 19 HOMOLOGUE. HDPCG79 691350 1060 WUblastx.64 (Q9H9H4) CDNA FLJ12750 FIS, Q9H9H4 98% 415 588 CLONE NT2RP2001168, WEAKLY SIMILAR TO VER HDPCV29 567228 1061 WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 40% 546 1196 human HDPDA36 827283 1062 WUblastx.64 (Q9CZU3) 2610528A15RIK Q9CZU3 96% 3443 3604 PROTEIN. HDPDC59 872451 1063 WUblastx.64 (Q9H251) CADHERIN RELATED Q9H251 99% 142 1737 23. 32% 145 1761 33% 130 1737 32% 130 1734 32% 112 1740 34% 145 1743 31% 124 1731 30% 145 1743 31% 256 1725 31% 145 1755 31% 145 1698 31% 139 1491 30% 658 1737 29% 154 1731 HDPFG13 637580 1064 WUblastx.64 (Q9H8S0) CDNA FLJ13287 FIS, Q9H8S0 100% 55 441 CLONE OVARC1001161. HDPFZ05 824390 1066 WUblastx.64 (Q9CZC8) 2810019K23RIK Q9CZC8 84% 500 658 PROTEIN. HDPGR80 844812 1068 WUblastx.64 (Q9BRR6) SIMILAR TO RIKEN Q9BRR6 87% 2434 1067 CDNA 2610017G09 GENE. 83% 2556 2377 HDPGU14 832543 1069 WUblastx.64 mda-9 protein - human pir|JC6537|JC6537 100% 292 1185 HDPGX09 580818 1070 WUblastx.64 (Q9D0M0) 2610002K22RIK Q9D0M0 94% 832 999 PROTEIN. 33% 995 1093 91% 1020 1220 HDPIE44 899328 1071 WUblastx.64 (Q9D666) 4632417G13RIK Q9D666 62% 102 2453 PROTEIN. HDPIE73 886158 1072 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 58% 1852 2145 CLONE COL04765. HDPIF35 826781 1073 WUblastx.64 (BAB55422) CDNA FLJ14966 fis, BAB55422 100% 1000 887 clone THYRO1000034, w HDPIF65 847047 1074 WUblastx.64 (Q9H7Z0) CDNA FLJ14058 FIS, Q9H7Z0 60% 236 12 CLONE HEMBB1000554. HDPIH25 740749 1075 WUblastx.64 (Q9H7Q7) FLJ00010 PROTEIN Q9H7Q7 78% 99 539 (FRAGMENT). 100% 876 968 76% 971 2266 98% 536 796 50% 2190 2300 75% 796 876 HDPIY31 886159 1076 WUblastx.64 hypothetical protein pir|T46448|T46448 72% 1714 1899 DKFZp434N1429.1 - human (fragment) HDPJH72 847048 1077 WUblastx.64 (O15427) MONOCARBOXYLATE MOT4_HUMAN 96% 294 392 TRANSPORTER 4 (MCT 4) (MCT 3). 81% 1204 1473 100% 77 295 89% 854 1192 37% 1427 1498 71% 394 855 HDPKC55 822869 1080 HMMER PFAM: Tubulin/FtsZ family PF00091 177.8 108 797 2.1.1 WUblastx.64 (Q9UJT1) TUBULIN DELTA CHAIN TBD_HUMAN 100% 102 1460 (DELTA TUBULIN). HDPKD16 897304 1081 WUblastx.64 (AAG10506) 2P domain K+ channel AAG10506 100% 246 857 TWIK-2. HDPMC52 847049 1082 WUblastx.64 (Q9Y275) TUMOR NECROSIS T13B_HUMAN 98% 67 447 FACTOR LIGAND SUPERFAMILY 100% 7 72 MEMBER 13 HDPML04 822851 1083 WUblastx.64 X-linked retinopathy protein (C- pir|A46010|A46010 73% 1403 1326 terminal, clone XEH.8c) - human 68% 1327 1223 (fragment) HDPNC21 839263 1085 WUblastx.64 (AAK55521) PRO0764. AAK55521 81% 1290 964 HDPNJ26 852338 1086 WUblastx.64 (Q9CQ35) 4921508O11RIK Q9CQ35 67% 115 651 PROTEIN. HDPOD73 876202 1087 WUblastx.64 (Q9P195) PRO1722. Q9P195 84% 173 211 60% 16 90 61% 78 185 HDPOT33 892319 1088 WUblastx.64 (Q92482) AQUAPORIN 3. AQP3_HUMAN 100% 53 928 HDPPB70 874027 1089 WUblastx.64 (Q96DV8) Unknown (protein for Q96DV8 68% 257 1186 MGC: 3551). HDPPE05 809109 1091 WUblastx.64 (Q9P0G1) HSPC075 (FRAGMENT). Q9P0G1 100% 355 459 100% 64 153 HDPSA70 722216 1092 WUblastx.64 (Q9UL01) SQUAMOUS CELL Q9UL01 100% 13 261 CARCINOMA ANTIGEN RECOGNIZED BY T CELL. HDPTI49 847052 1097 WUblastx.64 (Q14185) DOCK180 PROTEIN. Q14185 79% 927 1190 HDPYE25 636058 1099 WUblastx.64 (O42346) NEURULA-SPECIFIC O42346 73% 3 722 FERRODOXIN REDUCTASE-LIKE PROTEIN. HDQGD06 838076 1100 WUblastx.64 (Q9H1C4) UNC-93 RELATED Q9H1C4 80% 969 1535 PROTEIN. 90% 59 463 87% 454 996 HDQGD06 852673 1101 WUblastx.64 (Q9H1C4) UNC-93 RELATED Q9H1C4 81% 969 1559 PROTEIN. 90% 59 463 87% 454 996 HDQGD06 881280 1102 WUblastx.64 (Q9H1C4) UNC-93 RELATED Q9H1C4 86% 59 1537 PROTEIN. HDQGN08 840588 1103 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 56% 782 510 CLONE HEMBA1006036. HDQGO62 837353 1104 WUblastx.64 (Q9D6E9) 2900070E19RIK Q9D6E9 96% 222 515 PROTEIN. HDQPM16 886161 1105 WUblastx.64 (Q9DCP9) PROLINE RICH Q9DCP9 76% 145 648 PROTEIN EXPRESSED IN BRAIN. HDSAH37 603518 1109 WUblastx.64 (Q9H9U3) CDNA FLJ12547 FIS, Q9H9U3 41% 341 156 CLONE NT2RM4000634. 36% 449 147 35% 344 105 HDSAM57 834822 1110 WUblastx.64 (Q9P195) PRO1722. Q9P195 76% 752 690 80% 685 623 61% 897 742 HDSAO14 847054 1111 WUblastx.64 (Q9H0I1) HYPOTHETICAL 83.8 KDA Q9H0I1 100% 241 348 PROTEIN. HDSAP15 772712 1113 WUblastx.64 (Q9H059) HYPOTHETICAL 99.0 KDA Q9H059 46% 160 378 PROTEIN. HDTAR39 847055 1114 WUblastx.64 (Q9NVH2) HYPOTHETICAL 106.8 KDA Q9NVH2 100% 13 273 PROTEIN. 85% 242 772 HDTDA48 809110 1118 WUblastx.64 (O15254) PRISTANOYL-COA O15254 98% 149 601 OXIDASE. HDTDE66 610074 1119 WUblastx.64 (Q9Y609) LR8. Q9Y609 88% 1 447 HDTDG75 799892 1120 WUblastx.64 (Q9H2I8) CDA017. Q9H2I8 100% 434 514 HDTGW76 862012 1123 WUblastx.64 (Q9NY74) ETAA16 PROTEIN. Q9NY74 73% 1290 1835 27% 1422 1616 23% 296 829 82% 20 1234 HDTGZ56 800756 1124 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 54% 27 257 PROTEIN. HDTIM39 886164 1126 WUblastx.64 (Q9P0Q9) HSPC217. Q9P0Q9 80% 254 523 35% 302 403 90% 504 788 HDTKJ29 840023 1127 HMMER PFAM: Uncharacterized protein family PF01027 137.6 184 726 2.1.1 WUblastx.64 (Q9Y3C2) CGI-119 PROTEIN. Q9Y3C2 71% 19 732 HE2AC74 901343 1131 WUblastx.64 (AAK53385) G protein gamma 12 AAK53385 100% 207 422 subunit. HE2AX36 812337 1135 WUblastx.64 (Q9Y5B5) UBIQUITIN-SPECIFIC Q9Y5B5 85% 1292 300 PROTEASE HOMOLOG. HE2AY96 796800 1136 WUblastx.64 (BAB22850) 18 days embryo cDNA, BAB22850 87% 56 751 RIKEN full-length e HE2BD72 600349 1137 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 66% 496 371 CLONE KAIA0536. 77% 650 522 HE2BH50 847056 1138 WUblastx.64 (Q9GZW0) DJ604K5.1 (15 KDA Q9GZW0 100% 226 405 SELENOPROTEIN). HE2CC17 566806 1140 WUblastx.64 (Q9HBW6) NAG13. Q9HBW6 69% 996 694 59% 731 516 75% 347 180 42% 1037 933 70% 540 352 HE2CJ53 637592 1141 WUblastx.64 (Q9NUM3) CDNA FLJ11274 FIS, Q9NUM3 100% 123 266 CLONE PLACE1009368, WEAKLY 100% 266 412 SIMILAR TO MET HE2DI16 604029 1145 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 66% 1350 1201 CLONE KAIA0536. 76% 1483 1331 HE2DJ84 746643 1146 WUblastx.64 (P82673) MITOCHONDRIAL 28S P82673 100% 2 313 RIBOSOMAL PROTEIN S28 (MRP- S28). HE2DY23 877491 1147 WUblastx.64 (Q9VW97) CG17149 PROTEIN. Q9VW97 48% 1018 1284 HE2EH45 581520 1150 WUblastx.64 (Q9H3F4) MSTP030. Q9H3F4 73% 690 815 63% 8 250 75% 97 735 HE2FE89 780314 1151 WUblastx.64 (Q9QYN1) ENH3. Q9QYN1 95% 254 541 HE2FR49 714456 1152 WUblastx.64 (Q9NUP7) CDNA FLJ11219 FIS, Q9NUP7 64% 63 104 CLONE PLACE1008122. 100% 92 400 HE2GB19 566808 1153 WUblastx.64 (O95245) ENVELOPE PROTEIN O95245 41% 397 519 (FRAGMENT). 39% 246 362 63% 140 247 HE2HB61 798113 1155 WUblastx.64 (Q9BS71) UNKNOWN (PROTEIN Q9BS71 100% 261 365 FOR MGC: 12301). HE2HF76 637594 1157 WUblastx.64 (Q9HAZ1) PROTEIN SERINE Q9HAZ1 100% 11 859 THREONINE KINASE CLK4. HE21E66 830436 1159 WUblastx.64 (AAH22050) Hypothetical 61.5 kDa AAH22050 98% 98 523 protein (Fragment) HE2NW57 638615 1160 WUblastx.64 (Q9UI58) PRO0483 PROTEIN. Q9UI58 57% 191 15 HE2OA95 637595 1161 WUblastx.64 (Q9UIK5) TMEFF2 PROTEIN Q9UIK5 99% 1647 1231 PRECURSOR (TRANSMEMBRANE PROTEIN TENB2) (TPEF HE2PB61 847379 1163 WUblastx.64 (Q9BWY7) DJ639P13.1 (NOVEL Q9BWY7 92% 3 719 PROTEIN SIMILAR TO RAT TRANSPORTER-LIKE PR HE2PI43 847380 1164 WUblastx.64 (Q9H8E8) CDNA FLJ13697 FIS, Q9H8E8 100% 234 407 CLONE PLACE2000172. HE6CJ41 604908 1166 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 54% 898 1050 PROTEIN. HE6EI30 847381 1169 WUblastx.64 (Q9BRG1) SIMILAR TO RIKEN Q9BRG1 100% 603 523 CDNA 1110020N13 GENE. HE8BP64 847059 1176 WUblastx.64 (Q9DBS0) HIPPOCAMPUS Q9DBS0 71% 10 1299 ABUNDANT GENE TRANSCRIPT 1. HE8BQ49 589443 1177 WUblastx.64 hypothetical protein - human pir|S72482|S72482 75% 343 474 transposon MER37 64% 105 248 HE8BR18 731933 1178 WUblastx.64 (Q9NW38) CDNA FLJ10335 FIS, Q9NW38 100% 654 1091 CLONE NT2RM2000669. HE8BT58 806144 1180 WUblastx.64 (Q9BX67) JUNCTIONAL Q9BX67 91% 80 937 ADHESION MOLECULE 3 PRECURSOR. HE8CC34 753266 1183 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 72% 2116 1829 CLONE COL04765. HE8DK52 658676 1186 WUblastx.64 (Q9D1U5) D730039F16RIK Q9D1U5 66% 778 813 PROTEIN. 64% 843 935 HE8DZ94 877630 1187 WUblastx.64 (Q9Y3C2) CGI-119 PROTEIN. Q9Y3C2 91% 1295 1657 HE8EX86 823176 1189 WUblastx.64 (AAK53401) Elongation factor G2. AAK53401 100% 1410 457 96% 2790 2593 88% 2579 1362 HE8FC10 693908 1190 HMMER PFAM: Zinc finger, C2H2 type PF00096 72.8 606 674 2.1.1 WUblastx.64 (Q9H8L4) CDNA FLJ13479 FIS, Q9H8L4 84% 300 731 CLONE PLACE1003738, WEAKLY 98% 6 173 SIMILAR TO ZIN 45% 438 731 45% 438 674 44% 438 674 39% 438 674 36% 438 674 40% 444 674 37% 438 674 37% 444 674 51% 6 134 48% 6 161 46% 6 134 46% 6 134 47% 9 134 48% 6 134 47% 9 134 40% 9 134 44% 48 134 HE8FG15 799520 1191 WUblastx.64 (O43491) PROTEIN 4.1-G. O43491 84% 226 1197 HE8FG24 733560 1192 WUblastx.64 (Q9HAQ1) GLYCEROL 3- Q9HAQ1 90% 11 76 PHOSPHATE PERMEASE. 86% 94 522 HE8FK78 852180 1193 WUblastx.64 (Q9H8A3) CDNA FLJ13837 FIS, Q9H8A3 100% 1 75 CLONE THYRO1000748, 80% 72 266 MODERATELY SIMILAR TO 88% 269 1372 HE8FR53 843807 1196 WUblastx.64 (Q9H373) PRO1107. Q9H373 98% 3411 3181 HE8MQ01 886800 1199 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 71% 2692 2483 HE8MS43 799521 1200 WUblastx.64 (Q9D7J4) 2310005N03RIK Q9D7J4 88% 1023 946 PROTEIN. 83% 1247 1155 77% 175 44 HE8NC81 862015 1202 WUblastx.64 (Q9H6B2) CDNA: FLJ22418 FIS, Q9H6B2 100% 74 106 CLONE HRC08590. 88% 168 923 HE8NC81 1096692 3106 HMMER PFAM: Immunoglobulin domain PF00047 28.3 563 814 2.1.1 WUblastx.64 (Q9H6B2) CDNA: FLJ22418 FIS, Q9H6B2 99% 419 1264 CLONE HRC08590. HE8QU21 838391 1204 WUblastx.64 (Q9BGZ4) HYPOTHETICAL 11.6 KDA Q9BGZ4 44% 1391 1125 PROTEIN. HE8SH74 833053 1205 WUblastx.64 (Q9H387) PRO2550. Q9H387 69% 2476 2441 50% 3129 3034 76% 3017 2868 HE8UX34 838170 1206 WUblastx.64 (Q9H8Q9) CDNA FLJ13310 FIS, Q9H8Q9 67% 7 162 CLONE OVARC1001453. HE9CI81 734918 1209 WUblastx.64 (Q9DB52) 2900009I07RIK Q9DB52 82% 2273 2016 PROTEIN. HE9CJ38 565082 1210 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 64% 1180 1079 PROTEIN. 54% 1380 1243 HE9CN58 562768 1212 WUblastx.64 (Q99710) YY1-ASSOCIATED Q99710 44% 6 353 FACTOR 2. HE9CV59 847061 1213 WUblastx.64 (Q9CYF7) 5730494N06RIK Q9CYF7 76% 73 408 PROTEIN. HE9DG54 821326 1214 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 53% 1508 1197 PRODUCT. HE9DH59 886059 1215 WUblastx.64 (AAK49519) Putative mitochondrial AAK49519 100% 54 419 solute carrier sp 94% 1534 2085 HE9EM54 560627 1218 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 57% 611 688 CLONE COL03536. 67% 756 902 HE9FH28 633737 1219 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 58% 965 591 CLONE COL04765. HE9HE13 868829 1220 WUblastx.64 (Q9P183) PRO2160. Q9P183 100% 434 183 HE9HE13 824328 1221 WUblastx.64 (Q9UI56) PROTEIN PRO0518. P518_HUMAN 100% 665 462 HE9HF59 664485 1222 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 45% 1593 1276 PROTEIN. HE9TA42 834972 1227 WUblastx.64 (P53992) PROTEIN TRANSPORT S24C_HUMAN 100% 1423 1674 PROTEIN SEC24C (SEC24- 98% 2099 2359 RELATED PR 85% 1674 2105 29% 1833 1943 38% 58 135 98% 8 1435 HEAAC21 581048 1228 WUblastx.64 (Q9NV24) CDNA FLJ10980 FIS, Q9NV24 100% 100 246 CLONE PLACE1001570. HEAAD63 603321 1231 WUblastx.64 (Q9H965) CDNA FLJ12983 FIS, Q9H965 84% 613 575 CLONE NT2RP3000002. 68% 789 613 HEAAM96 562769 1234 WUblastx.64 (Q9H4S5) DJ336K20B.1 (NOVEL Q9H4S5 83% 541 1044 PROTEIN BASED ON FGENESH) (FRAGMENT). HEAAN52 862020 1235 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 54% 796 1104 PROTEIN. HEAAW94 847340 1238 WUblastx.64 (Q9UEV9) ACTIN-BINDING Q9UEV9 94% 285 890 PROTEIN HOMOLOG ABP-278. 41% 285 884 38% 285 884 36% 285 872 35% 285 884 34% 285 884 34% 285 878 32% 303 887 33% 285 881 33% 285 878 31% 285 869 29% 288 887 30% 279 884 31% 285 863 36% 414 848 30% 288 884 32% 429 857 31% 309 824 26% 321 824 38% 285 539 37% 552 824 HEBAT05 637602 1240 WUblastx.64 (Q9NWW0) CDNA FLJ20568 FIS, Q9NWW0 96% 111 380 CLONE REC00775. 87% 84 131 HEBCN80 596808 1243 WUblastx.64 (Q9BGX7) HYPOTHETICAL 13.0 KDA Q9BGX7 57% 782 865 PROTEIN. 53% 666 788 HEEAF49 880521 1254 WUblastx.64 (Q9P147) PRO2822. Q9P147 79% 2117 1896 HEEAJ46 637623 1255 WUblastx.64 (AAK55521) PRO0764. AAK55521 86% 2358 2314 66% 1626 1582 55% 2336 2163 73% 1601 1368 HEGAI20 825818 1256 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 63% 893 759 PRODUCT. HEIAC52 826611 1257 WUblastx.64 (Q9H6A7) CDNA: FLJ22429 FIS, Q9H6A7 75% 40 915 CLONE HRC09084. HELAT58 732199 1259 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 56% 1807 1881 CLONE COL03536. 60% 1731 1805 59% 1880 2035 HELAW94 834796 1260 WUblastx.64 (AAH02585) RAB7, member RAS AAH02585 100% 159 551 oncogene family-like 1. HELDF80 799522 1261 WUblastx.64 N-ras upstream protein NRU - human pir|S29815|S29815 99% 3 1289 28% 6 1061 28% 435 1118 HELDK79 532597 1263 WUblastx.64 L6 surface protein - human pir|A42926|A42926 78% 71 676 HELDQ42 695716 1264 WUblastx.64 (O60629) BLADDER CANCER- BC10_HUMAN 100% 289 549 ASSOCIATED PROTEIN (BLADDER CANCER HELEL76 862490 1266 WUblastx.64 (Q9BRX8) SIMILAR TO RIKEN Q9BRX8 93% 171 818 CDNA 5730469M10 GENE. HELEL76 839470 1267 WUblastx.64 (Q9BRX8) SIMILAR TO RIKEN Q9BRX8 98% 545 694 CDNA 5730469M10 GENE. 90% 48 506 HELEO45 847067 1268 WUblastx.64 (Q9H6Q6) CDNA: FLJ21982 FIS, Q9H6Q6 92% 1385 1423 CLONE HEP06188. 88% 7 1392 HELFA57 845981 1269 WUblastx.64 (O95761) WUGSC: H_DJ0771P04.2 O95761 95% 340 1362 PROTEIN (FRAGMENT). 95% 9 137 78% 62 409 HELHN47 872795 1273 WUblastx.64 (Q9NWC8) HYPOTHETICAL 17.6 KDA Q9NWC8 91% 1233 772 PROTEIN. HELHP11 638064 1274 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 76% 1429 1313 CLONE COL03536. HELHP11 851116 1275 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 79% 1441 1313 CLONE COL03536. HEMBV40 847069 1277 WUblastx.64 NADH dehydrogenase (ubiquinone) pir|JC5822|JC5822 77% 798 1055 (EC 1.6.5.3) CI-B12 chain - human HEMCJ80 765041 1278 WUblastx.64 (Q9NXF8) CDNA FLJ20279 FIS, Q9NXF8 98% 489 710 CLONE HEP03229. 73% 3 206 79% 155 487 HEMDB07 617137 1280 WUblastx.64 (Q9H387) PRO2550. Q9H387 82% 400 660 HEMGK71 847374 1282 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 80% 1383 1258 CLONE KAIA0536. 64% 1576 1352 HEOMU44 778085 1287 WUblastx.64 (Q9N307) HYPOTHETICAL Q9N307 52% 823 1392 PROTEIN Y61A9LA.B. HEONI85 566833 1288 WUblastx.64 hypothetical 43.5K protein - mouse pir|JU0319|JU0319 77% 236 421 90% 382 672 HEONK04 658682 1289 WUblastx.64 (Q9UKN4) BRIDGING Q9UKN4 72% 267 1070 INTEGRATOR-2. HEONP08 601691 1290 WUblastx.64 (Q9Y349) HYPOTHETICAL 15.7 KDA Q9Y349 100% 924 523 PROTEIN. HEPBC23 799985 1292 WUblastx.64 (Q9H1V4) DJ1182A14.3 (SIMILAR Q9H1V4 92% 4 303 TO MST1 (MACROPHAGE STIMULATING 1 (HEPA HEPCU48 805585 1295 WUblastx.64 (Q9UNT2) PROTEIN O- Q9UNT2 79% 315 1490 MANNOSYL-TRANSFERASE 1. 95% 79 324 100% 15 32 HEQAH47 877634 1296 WUblastx.64 hypothetical protein pir|T08772|T08772 100% 630 1085 DKFZp586M121.1 - human 87% 12 476 (fragment) 30% 90 308 30% 189 305 29% 1104 1355 28% 30 245 63% 257 289 HEQBJ01 861786 1300 WUblastx.64 (Q9LVQ7) ZINC FINGER PROTEIN. Q9LVQ7 34% 424 849 HEQBJ01 876546 1301 WUblastx.64 (Q9LVQ7) ZINC FINGER PROTEIN. Q9LVQ7 34% 424 849 HEQBM94 580823 1302 WUblastx.64 (Q9BT25) UNKNOWN (PROTEIN Q9BT25 100% 470 559 FOR IMAGE: 3636299) 72% 583 843 (FRAGMENT). 77% 7 471 HESAG57 855936 1307 WUblastx.64 (O43432) EIF4GII. O43432 82% 936 7 100% 1221 1030 100% 1365 1336 27% 1382 1254 28% 1545 1327 HETBJ88 855937 1310 WUblastx.64 (Q9H1Y3) DJ317G22.2 Q9H1Y3 83% 10 864 (ENCEPHALOPSIN) (PANOPSIN). HETCM67 861909 1311 HMMER PFAM: Disintegrin PF00200 112.2 239 466 2.1.1 WUblastx.64 (Q9UKQ2) ADAM 28 PRECURSOR AD28_HUMAN 96% 11 1318 (EC 3.4.24.—) (A DISINTEGRIN AND HETDD61 800668 1312 WUblastx.64 (Q9Y6B7) ADAPTER-RELATED A4B1_HUMAN 96% 789 1085 PROTEIN COMPLEX 4 BETA 1 85% 363 569 SUBUNIT ( 96% 1315 1761 HETDD61 852381 1313 WUblastx.64 (Q9Y6B7) ADAPTER-RELATED A4B1_HUMAN 96% 1314 1760 PROTEIN COMPLEX 4 BETA 1 96% 789 1085 SUBUNIT ( 85% 363 569 HETDP76 782836 1316 WUblastx.64 (Q9BUK6) HYPOTHETICAL 61.8 KDA Q9BUK6 100% 29 394 PROTEIN. 92% 700 1620 80% 1895 2116 53% 1428 1466 100% 465 653 HETFO57 844190 1317 WUblastx.64 (Q9H229) RAD50-INTERACTING Q9H229 96% 2 1363 PROTEIN 1. HFAAI17 607563 1322 WUblastx.64 (Q9H387) PRO2550. Q9H387 61% 426 518 73% 281 403 HFADF41 607557 1324 WUblastx.64 B-cell growth factor precursor - human pir|A47582|A47582 56% 600 737 HFADM09 899435 1325 WUblastx.64 ISOFORM 8 OF Q9Y4J8 sp_vs|Q9Y4J8- 100% 382 480 02|Q9Y4J8 HFAUA23 636071 1326 WUblastx.64 (Q9HAL3) CDNA FLJ11393 FIS, Q9HAL3 25% 243 91 CLONE HEMBA1000591, WEAKLY 45% 466 263 SIMILAR TO PTB HFCAG75 834834 1327 WUblastx.64 (AAK55521) PRO0764. AAK55521 80% 745 701 78% 720 484 HFCAI40 866441 1328 HMMER PFAM: TPR Domain PF00515 52.6 167 253 2.1.1 WUblastx.64 (Q9V532) CG8777 PROTEIN. Q9V532 36% 20 1021 HFCAQ17 695720 1329 WUblastx.64 NADH dehydrogenase (ubiquinone) pir|JE0379|JE0379 78% 20 238 (EC 1.6.5.3) chain NDUFA3 - human HFCBC16 559312 1330 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 55% 998 792 CLONE COL03536. HFCBT29 654842 1334 WUblastx.64 (Q9H9H1) CDNA FLJ12756 FIS, Q9H9H1 73% 80 280 CLONE NT2RP2001295, WEAKLY SIMILAR TO ZIN HFCDN13 722221 1336 WUblastx.64 (Q9P0T7) HSPC186 Q9P0T7 90% 2 187 (HYPOTHETICAL 20.6 KDA PROTEIN). HFCDT67 598900 1337 WUblastx.64 (Q9H0N3) HYPOTHETICAL 107.1 KDA Q9H0N3 82% 9 455 PROTEIN. 77% 845 910 45% 373 438 67% 498 902 HFCDY36 834512 1338 WUblastx.64 catalase (EC 1.11.1.6) - pir|I40767|I40767 93% 279 190 Campylobacter jejuni HFCEC45 847384 1339 WUblastx.64 (Q9D3Q4) 5033428A16RIK Q9D3Q4 98% 68 349 PROTEIN. HFCET43 596813 1340 WUblastx.64 (Q9Z1M7) Q9Z1M7 100% 7 144 ACETYLGLUCOSAMINYLTRANSFERASE- LIKE PROTEIN. HFEAG55 838142 1341 WUblastx.64 (Q9EP97) SENTRIN/SUMO- Q9EP97 100% 12 200 SPECIFIC PROTEASE (SMT3 ISOPEPTIDASE 1). HFEAU63 790190 1342 WUblastx.64 ISOFORM EYA1B OF Q99502 sp_vs|Q99502- 90% 2180 2028 01|Q99502 HFEBA88 684297 1343 WUblastx.64 (BAB55210) CDNA FLJ14675 fis, BAB55210 100% 205 297 clone NT2RP2003952, w 44% 70 327 HFEBO15 831772 1345 WUblastx.64 (Q9HC46) HYPOTHETICAL 42.8 KDA Q9HC46 85% 226 477 PROTEIN. 53% 150 332 51% 2 298 HFEBO17 852218 1346 WUblastx.64 (BAB55130) CDNA FLJ14559 fis, BAB55130 100% 523 624 clone NT2RM2001998. 91% 606 809 HFFAE46 654843 1347 WUblastx.64 (O14597) NON-FUNCTIONAL O14597 100% 608 658 FOLATE BINDING PROTEIN. 96% 703 801 78% 386 442 58% 602 649 57% 402 632 HFFAH01 843438 1348 WUblastx.64 hypothetical protein pir|T43460|T43460 91% 510 241 DKFZp434P1721.1 - human 98% 259 11 (fragment) 95% 818 513 HFFAL70 827570 1349 WUblastx.64 (AAH07384) Unknown (protein for AAH07384 84% 921 466 IMAGE: 3677194) (Fra HFFAV61 664486 1350 WUblastx.64 (BAA92711) Adaptor protein BAA92711 100% 1555 1184 p130Cas. 50% 577 536 HFGAN63 708053 1353 WUblastx.64 (Q9D4E9) 4932701A20RIK Q9D4E9 34% 159 542 PROTEIN. 52% 561 617 HFIAB78 581054 1355 WUblastx.64 (Q9UKA1) F-BOX PROTEIN FBL5 Q9UKA1 100% 672 872 (FRAGMENT). 100% 14 667 58% 608 658 32% 461 661 36% 690 836 HFIIK29 843587 1360 WUblastx.64 (Q9H705) CDNA: FLJ21611 FIS, Q9H705 94% 48 218 CLONE COL07344. HFIIK29 852308 1361 WUblastx.64 (Q9H705) CDNA: FLJ21611 FIS, Q9H705 94% 48 218 CLONE COL07344. HFIIK29 873681 1362 WUblastx.64 (Q9H705) CDNA: FLJ21611 FIS, Q9H705 94% 48 218 CLONE COL07344. HFIIK29 885007 1363 WUblastx.64 (Q9H705) CDNA: FLJ21611 FIS, Q9H705 94% 48 218 CLONE COL07344. HFIIZ61 855939 1366 WUblastx.64 (Q62786) PROSTAGLANDIN F2- FPRP_RAT 92% 35 229 ALPHA RECEPTOR REGULATORY PROTEIN HFIUT21 794323 1372 WUblastx.64 (AAH07934) DKFZP434A043 AAH07934 100% 6 320 protein. HFIXC39 683053 1374 WUblastx.64 hypothetical protein 1 - rat (fragment) pir|S33477|S33477 36% 1816 1541 HFIXC69 704080 1375 WUblastx.64 (Q9NRX3) NADH: UBIQUINONE Q9NRX3 87% 32 292 OXIDOREDUCTASE MLRQ SUBUNIT HOMOLOG. HFIXE39 844408 1376 WUblastx.64 (CAC18649) Lipoyl-containing CAC18649 100% 1394 1651 component X precursor. HFIYP15 688044 1377 WUblastx.64 (Q9H324) ZINC Q9H324 49% 430 1074 METALLOENDOPEPTIDASE 35% 406 915 (FRAGMENT). 31% 553 1110 36% 712 966 33% 577 738 34% 481 558 HFKBC47 847076 1383 WUblastx.64 (O95302) FK506-BINDING O95302 99% 1177 323 PROTEIN (FRAGMENT). 54% 1174 404 47% 997 323 HFKDX53 731867 1384 WUblastx.64 (Q9BTV6) UNKNOWN (PROTEIN Q9BTV6 100% 407 763 FOR IMAGE: 3502107) 65% 204 371 (FRAGMENT). 100% 766 1296 HFKEB14 709663 1385 WUblastx.64 (Q9NVY4) CDNA FLJ10436 FIS, Q9NVY4 97% 8 127 CLONE NT2RP1000574, HIGHLY SIMILAR TO HOM HFKEU17 815554 1389 WUblastx.64 (AAH00465) Growth arrest and DNA- AAH00465 97% 226 330 damage-inducible, 86% 473 784 HFKEV77 850678 1390 WUblastx.64 hypothetical protein pir|T14797|T14797 89% 238 567 DKFZp564B167.1 - human HFKFI15 775846 1391 WUblastx.64 (Q12948) FORKHEAD BOX FXC1_HUMAN 100% 2 166 PROTEIN C1 (FORKHEAD- RELATED PROTEIN HFKFK49 847077 1393 WUblastx.64 (Q9UHR0) MAP KINASE- Q9UHR0 100% 277 366 INTERACTING KINASE. HFOXD49 844524 1398 WUblastx.64 (O15194) YA22 PROTEIN (HYA22). O15194 99% 7 435 HFOXE28 847385 1399 WUblastx.64 (Q9H0E0) HYPOTHETICAL 13.4 KDA Q9H0E0 47% 1011 1286 PROTEIN. 39% 135 476 34% 360 671 60% 837 881 HFOYR54 743196 1403 WUblastx.64 (O00566) U3 SMALL NUCLEOLAR MP10_HUMAN 78% 118 1365 RIBONUCLEOPROTEIN PROTEIN 63% 136 168 MPP10 HFPBQ55 608171 1409 WUblastx.64 (Q24333) ELASTIN LIKE PROTEIN Q24333 100% 45 116 (FRAGMENT). HFPCM32 608215 1411 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 48% 719 441 CLONE COL03536. HFPCU47 741049 1414 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 59% 1645 1580 CLONE COL04765. 58% 1598 1356 HFPCY66 825718 1415 WUblastx.64 (Q9H387) PRO2550. Q9H387 72% 1371 1252 72% 1555 1424 HFPDC65 839208 1416 WUblastx.64 (Q9H5G8) CDNA: FLJ23451 FIS, Q9H5G8 100% 72 605 CLONE HSI06456. HFPDE42 844402 1417 WUblastx.64 (Q99PQ3) TRIPARTITE MOTIF Q99PQ3 97% 3 218 PROTEIN TRIM9 (FRAGMENT). HFPDP70 745433 1420 WUblastx.64 (Q9H919) CDNA FLJ13078 FIS, Q9H919 83% 937 990 CLONE NT2RP3002002. 88% 879 905 58% 980 1096 HFPDX08 746379 1422 WUblastx.64 (Q9H9Y3) CDNA FLJ12474 FIS, Q9H9Y3 100% 3 311 CLONE NT2RM1000927. HFPEP69 835459 1423 WUblastx.64 (Q9P0X1) LIPOPOLYSACCHARIDE Q9P0X1 96% 654 749 SPECIFIC RESPONSE-5 PROTEIN (FRAGMENT). HFRAU40 701986 1424 WUblastx.64 hypothetical protein 3 - human pir|E41925|E41925 34% 228 109 57% 511 428 45% 428 252 HFSAY91 828067 1426 WUblastx.64 hypothetical protein pir|T46933|T46933 100% 1280 1197 DKFZp434M035.1 - human 92% 1096 437 HFSBC10 638315 1427 WUblastx.64 (O95025) SEMAPHORIN 3D SM3D_HUMAN 82% 1437 532 PRECURSOR. 82% 626 231 54% 742 443 100% 234 67 HFSBE94 600398 1428 WUblastx.64 (Q9P195) PRO1722. Q9P195 75% 705 610 73% 853 695 HFTAN11 638316 1429 WUblastx.64 (Q9H387) PRO2550. Q9H387 54% 676 614 82% 771 667 68% 537 490 83% 617 489 HFTBL17 844488 1434 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 95% 967 710 PROTEIN (FRAGMENT). HFTBL17 863152 1435 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 95% 969 712 PROTEIN (FRAGMENT). HFTCI85 664487 1437 WUblastx.64 (O60448) NEURONAL THREAD O60448 63% 1140 1075 PROTEIN AD7C-NTP. 92% 1070 1032 38% 1139 1032 67% 1285 1130 59% 1067 1002 33% 877 707 65% 1275 1129 77% 1283 1257 72% 1276 1019 HFTCJ32 604911 1438 WUblastx.64 (Q9H387) PRO2550. Q9H387 59% 669 586 71% 889 689 HFTCO17 844354 1439 WUblastx.64 (Q9QY01) SERINE/THREONINE Q9QY01 83% 1 231 KINASE UNC51.2. 95% 216 611 HFTCW07 604987 1440 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 75% 798 589 CLONE LNG09295. HFTDF32 600360 1441 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 73% 1017 1310 CLONE COL04765. HFTDF79 827459 1442 WUblastx.64 hypothetical protein XF0051 pir|G82853|G82853 40% 873 691 [imported] - Xylella fastidiosa (strain 32% 872 627 9a5c) 51% 1202 1113 42% 813 589 32% 961 716 28% 792 424 62% 1184 1113 62% 1184 1113 42% 873 619 HFTDK11 837382 1443 WUblastx.64 (Q9Y6D7) ES18 (FRAGMENT). Q9Y6D7 100% 2 67 100% 558 650 63% 97 618 HFTDU08 825715 1444 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 70% 885 775 CLONE KAIA0536. HFVGK67 569778 1445 WUblastx.64 (Q9H387) PRO2550. Q9H387 83% 1269 985 HFVHD38 826709 1446 WUblastx.64 SHORT ISOFORM OF O00339 sp_vs|O00339- 98% 2163 550 01|O00339 56% 2163 1387 50% 2163 1387 49% 2163 1387 48% 2142 1387 40% 1386 820 HFVIC33 799524 1448 HMMER PFAM: Enoyl-CoA PF00378 34.7 462 680 2.1.1 hydratase/isomerase family WUblastx.64 (Q9NTX5) DJ351K20.2.1 (NOVEL Q9NTX5 96% 694 975 ENOYL COA/ACYL COA 99% 72 716 HYDRATASE/DEHYDROGENA HFXAK32 638319 1449 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 40% 1403 1104 PROTEIN. HFXAK59 831141 1450 WUblastx.64 (Q9BTC8) SIMILAR TO Q9BTC8 95% 1537 431 METASTASIS ASSOCIATED 1. HFXBM52 737672 1453 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 80% 708 631 CLONE MAMMA1000472. 50% 123 64 68% 569 465 HFXBV67 748230 1455 WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 52% 1618 1319 human HFXCI42 606806 1458 WUblastx.64 (Q9GMI7) HYPOTHETICAL 9.0 KDA Q9GMI7 43% 542 297 PROTEIN. 72% 534 406 HFXCL59 825713 1459 WUblastx.64 (Q9NSI6) WD-REPEAT PROTEIN 9 WDR9_HUMAN 90% 1008 397 (FRAGMENT). 76% 53 3 35% 1002 385 28% 993 604 24% 960 556 29% 349 182 100% 367 182 88% 180 154 HFXCS53 828903 1462 WUblastx.64 (Q9H8H0) CDNA FLJ13640 FIS, Q9H8H0 99% 1107 442 CLONE PLACE1011221. HFXDB37 569805 1463 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 58% 1116 1208 PROTEIN. 57% 991 1131 HFXDL76 821571 1466 WUblastx.64 probable pol polyprotein-related pir|S21348|S21348 63% 1811 1746 protein 4 - rat 48% 1520 1446 64% 1740 1519 HFXDP44 590269 1469 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 64% 531 761 CLONE KAT08285. 50% 918 1115 88% 745 825 36% 1005 1103 30% 660 779 HFXDR08 621342 1470 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 64% 506 700 PROTEIN. HFXDR28 621300 1471 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 68% 652 717 CLONE LNG09295. 70% 501 653 HFXDR28 873859 1472 WUblastx.64 (Q9H5X2) CDNA: FLJ22865 FIS, Q9H5X2 98% 853 314 CLONE KAT02171. HFXDR47 846477 1473 WUblastx.64 (Q9H387) PRO2550. Q9H387 100% 977 957 92% 1051 1010 65% 957 733 HFXDZ03 846323 1474 WUblastx.64 (Q9UN78) HYPOTHETICAL 40.1 KDA Q9UN78 32% 1065 739 PROTEIN. 52% 528 385 24% 1161 877 39% 757 521 HFXEE88 838822 1476 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 56% 1310 1116 CLONE KAT08285. HFXGR32 534607 1477 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 57% 24 272 CLONE MAMMA1000472. HFXGT51 638174 1478 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 74% 1126 989 CLONE KAIA0536. 78% 1232 1107 HFXHI33 874255 1481 WUblastx.64 (Q9HBS7) HYPOTHETICAL 14.2 KDA Q9HBS7 66% 1535 1278 PROTEIN. HFXHL21 581363 1482 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 56% 1441 1373 PROTEIN. 64% 1601 1443 HFXHM93 858720 1485 WUblastx.64 (AAK55521) PRO0764. AAK55521 66% 803 1027 HFXHN89 786196 1486 WUblastx.64 (O60448) NEURONAL THREAD O60448 61% 1063 1218 PROTEIN AD7C-NTP. 59% 1208 1273 65% 1072 1332 HFXJB21 743165 1487 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 73% 1269 1003 CLONE COL04765. HFXJN93 596815 1488 WUblastx.64 (Q9D4B1) 4933405A16RIK Q9D4B1 97% 501 298 PROTEIN. HFXJS15 800866 1489 WUblastx.64 (Q9ESN6) NEURAL ACTIVITY- Q9ESN6 97% 1 447 RELATED RING FINGER PROTEIN 32% 37 438 (TRIPARTITE MOTI 33% 10 435 31% 10 516 HFXJT53 823591 1490 WUblastx.64 (AAL50053) Viperin. AAL50053 94% 111 1193 HFXKL60 895307 1492 WUblastx.64 (Q9H3C0) PRO0898. Q9H3C0 37% 1749 1889 67% 1865 2029 HFXLG08 885514 1493 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 60% 1303 1067 PROTEIN. HFXLK91 827288 1494 WUblastx.64 (Q9NWT2) CDNA FLJ20623 FIS, Q9NWT2 87% 3 827 CLONE KAT04793. HGBBR29 823106 1497 WUblastx.64 (O43592) EXPORTIN T. O43592 100% 183 1118 HGBDV35 581058 1499 WUblastx.64 (AAK53702) ADMP. AAK53702 42% 91 252 HGBHE23 832303 1502 WUblastx.64 hypothetical protein pir|T12479|T12479 86% 24 455 DKFZp564N1362.1 - human (fragment) HGBHI15 608170 1503 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 63% 663 565 PROTEIN. 65% 559 380 HGLAG32 668271 1505 WUblastx.64 (Q9BRZ3) UNKNOWN (PROTEIN Q9BRZ3 66% 2 154 FOR MGC: 2724). HGLAH86 603525 1507 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 55% 824 618 CLONE KAT08285. HHEBP28 775744 1517 WUblastx.64 (Q9BRL7) SIMILAR TO VESICLE Q9BRL7 100% 3 233 TRAFFICKING PROTEIN. HHECR10 621371 1519 WUblastx.64 (Q9H7S6) CDNA FLJ14310 FIS, Q9H7S6 64% 972 766 CLONE PLACE3000271. HHEMC55 566796 1520 WUblastx.64 (O60448) NEURONAL THREAD O60448 64% 605 417 PROTEIN AD7C-NTP. 42% 659 417 63% 1063 896 48% 912 808 41% 575 405 47% 1063 965 40% 1017 781 42% 976 878 61% 1060 827 32% 1063 812 50% 896 771 80% 349 320 65% 656 597 40% 606 325 43% 516 343 45% 941 885 65% 606 445 HHEMP35 821328 1523 WUblastx.64 (Q9Y6X2) PROTEIN INHIBITOR OF Q9Y6X2 93% 302 931 ACTIVATIED STAT3 (PROTEIN 60% 75 308 INHIBITOR OF HHEMZ08 840026 1524 WUblastx.64 (Q9NZ80) UNCHARACTERIZED Q9NZ80 77% 845 1351 BONE MARROW PROTEIN BM042. HHENR74 825671 1527 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 70% 1005 1085 CLONE COL04765. 77% 811 876 76% 878 1003 HHENY07 638321 1529 WUblastx.64 (O00311) CELL DIVISION CYCLE CDC7_HUMAN 88% 8 136 7-RELATED PROTEIN KINASE (EC 2 98% 121 744 HHEOK77 841099 1530 WUblastx.64 (Q9Z1W6) P80 (FRAGMENT). Q9Z1W6 76% 238 276 87% 942 1127 23% 510 764 80% 315 977 HHEPE72 621254 1531 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 53% 1015 749 CLONE COL04765. HHEPE81 840614 1532 WUblastx.64 (O95373) RANBP7/IMPORTIN 7. O95373 86% 3 389 HHEQY60 799535 1535 WUblastx.64 (O60448) NEURONAL THREAD O60448 68% 703 566 PROTEIN AD7C-NTP. 62% 731 555 68% 568 503 47% 570 502 58% 489 439 65% 731 429 HHFEB79 1300768 1541 WUblastx.64 (Q92545) RW1 PROTEIN RW1_HUMAN 74% 708 2387 (FRAGMENT). 80% 6 590 32% 1984 2148 HHFEB79 863749 3107 WUblastx.64 Similar to a C. elegans protein encoded dbj|BAA13387.1| 69% 1303 2400 in cosmid C27F2 (U40419) [Homo 80% 601 1185 sapiens] HHFEN34 701992 1543 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 63% 709 644 CLONE HEMBA1006036. 57% 942 691 HHFFZ01 847388 1544 WUblastx.64 (Q9H6Y7) CDNA: FLJ21676 FIS, Q9H6Y7 89% 835 1080 CLONE COL09164. 91% 1077 1763 HHFGI71 663531 1545 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 75% 1174 1272 PROTEIN. 67% 1424 1269 HHFGJ54 731870 1546 WUblastx.64 (Q9H906) CDNA FLJ13099 FIS, Q9H906 99% 1232 1645 CLONE NT2RP3002248. 96% 510 1259 HHFGL38 833063 1547 WUblastx.64 (Q9H387) PRO2550. Q9H387 53% 2764 2465 HHFGZ23 721653 1549 WUblastx.64 hypothetical protein 3 - human pir|E41925|E41925 64% 978 1136 HHFHM47 638181 1551 WUblastx.64 (O60448) NEURONAL THREAD O60448 76% 680 618 PROTEIN AD7C-NTP. 40% 804 670 65% 802 671 84% 611 573 72% 681 616 60% 77 3 62% 838 557 HHGAA76 767745 1552 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 49% 1301 870 PROTEIN (FRAGMENT). 69% 499 431 68% 59 3 74% 1455 1294 66% 888 505 HHGAD46 812646 1553 WUblastx.64 (Q9UGT4) BK65A6.2 (NOVEL Q9UGT4 85% 148 783 SUSHI DOMAIN (SCR REPEAT) 67% 3 203 CONTAINING PROTEIN HHGBC21 840455 1555 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 54% 1118 858 CLONE COL03536. HHGBW55 638183 1559 WUblastx.64 hypothetical protein [imported] - pir|T50835|T50835 97% 3 143 human HHGDR05 610266 1564 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 63% 821 690 CLONE HEMBA1006036. HHGDR92 840454 1565 WUblastx.64 (Q9Y512) PROTEIN CGI-51. CG51_HUMAN 99% 10 1104 HHGDS56 663379 1566 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 57% 1318 1187 CLONE KAIA0536. 62% 1476 1309 HHGDW65 825652 1567 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 56% 463 726 PROTEIN. HHPDE28 877640 1571 WUblastx.64 (Q9H620) CDNA: FLJ22673 FIS, Q9H620 98% 393 596 CLONE HSI10503. HHPEB61 658693 1575 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 50% 777 935 CLONE COL03536. 54% 622 804 HHPFP26 753269 1576 WUblastx.64 (Q9BQG8) HYPOTHETICAL 32.5 KDA Q9BQG8 100% 906 1742 PROTEIN. HHPFS11 570807 1577 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 50% 402 587 CLONE COL04765. HHPFS18 829347 1579 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 75% 2558 2427 CLONE COL04765. HHPGH34 535780 1580 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 68% 1019 972 FOR MGC: 5149). 64% 835 794 68% 963 850 HHPGU87 658694 1582 WUblastx.64 SHORT ISOFORM OF O60658 sp_vs|O60658- 100% 1050 1274 01|O60658 HHPSE55 735562 1585 WUblastx.64 (Q9CQ15) 0610011N22RIK Q9CQ15 79% 67 846 PROTEIN (RIKEN CDNA 0610011N22 GENE). HHPSF70 709665 1586 WUblastx.64 (AAH18471) Similar to nitrogen AAH18471 81% 1 96 fixation gene 1 (S. HHPSH74 638185 1587 WUblastx.64 hypothetical protein DKFZp566E044.1 - pir|T46284|T46284 72% 323 1339 human 99% 10 321 54% 10 321 53% 323 571 20% 752 1030 87% 1272 1730 HHPTF26 857654 1590 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 66% 1006 752 PROTEIN. HHSAE74 839473 1592 WUblastx.64 (Q9P1J1) PRO1546. Q9P1J1 89% 451 365 87% 588 442 HHSAG62 847392 1593 WUblastx.64 (Q9H876) CDNA FLJ13898 FIS, Q9H876 99% 2 460 CLONE THYRO1001738, WEAKLY 99% 600 1052 SIMILAR TO TUB 100% 533 604 HHSBJ92 838604 1595 WUblastx.64 (Q9BTM2) SIMILAR TO RIKEN Q9BTM2 100% 509 718 CDNA 1300006O23 GENE (FRAGMENT). HHSDB43 812510 1600 HMMER PFAM: Zinc finger, C2H2 type PF00096 117.4 877 945 2.1.1 WUblastx.64 hypothetical protein pir|T42688|T42688 92% 25 1344 DKFZp434H0127.1 - human (fragment) HHSDL07 899430 1601 WUblastx.64 (Q9UJH9) C380A1.2.1 (NOVEL Q9UJH9 85% 9 317 PROTEIN (ISOFORM 1)) (UNKNOWN) (PROTEIN FO HHSDX07 847393 1602 WUblastx.64 (Q9H9W2) CDNA FLJ12510 FIS, Q9H9W2 86% 3 512 CLONE NT2RM2001723. 79% 434 1066 HHSFF54 836149 1603 WUblastx.64 (Q9BS18) UNKNOWN (PROTEIN Q9BS18 100% 129 350 FOR MGC: 12537). HIABC70 838605 1607 WUblastx.64 (Q9NPJ9) APOLIPOPROTEIN B48 Q9NPJ9 77% 15 851 RECEPTOR. 100% 1047 1160 34% 522 821 25% 303 842 24% 306 818 24% 312 839 HIBCO70 853360 1609 HMMER PFAM: Myelin proteolipid protein PF01275 32.3 127 192 2.1.1 (PLP or lipophilin) WUblastx.64 (Q99NH0) GENE TRAP ANKYRIN Q99NH0 95% 176 1564 REPEAT CONTAINING PROTEIN. 38% 410 1414 39% 413 1408 37% 419 1411 39% 413 1408 38% 410 1360 37% 443 1441 38% 425 1420 37% 608 1411 38% 407 1141 33% 887 1411 HIBDA41 603909 1611 WUblastx.64 (Q9H387) PRO2550. Q9H387 80% 404 697 HIBEC45 822854 1612 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 63% 1023 1283 PROTEIN. HILBW03 841377 1613 WUblastx.64 (Q9NVS4) CDNA FLJ10546 FIS, Q9NVS4 97% 705 1163 CLONE NT2RP2001721. HISAE16 845233 1614 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 85% 7 90 CLONE COL04765. HISAG53 843902 1615 WUblastx.64 (BAB55426) CDNA FLJ14972 fis, BAB55426 100% 84 122 clone THYRO1000715. 83% 952 1614 78% 127 993 HISAN63 797658 1616 WUblastx.64 (O00363) PUTATIVE P150. O00363 33% 1609 1427 29% 1299 1045 59% 1454 1392 35% 716 384 HISAT78 840961 1617 WUblastx.64 (Q9NUD5) DJ1103G7.7 (PUTATIVE Q9NUD5 66% 155 280 NOVEL PROTEIN). 96% 13 108 29% 594 767 100% 114 194 32% 591 785 50% 538 567 100% 576 743 98% 280 567 HISBA38 561711 1618 WUblastx.64 (Q9H387) PRO2550. Q9H387 53% 919 836 53% 996 907 51% 842 687 HISBB66 839808 1619 WUblastx.64 (O95627) POTENTIAL O95627 88% 2 451 TRANSCRIPTIONAL REPRESSOR NOT4HP. HISEJ52 886777 1623 WUblastx.64 (Q9Y6J0) CALCINEURIN-BINDING CABI_HUMAN 94% 480 1697 PROTEIN CABIN 1 (CALCINEURIN I HJABC58 753272 1624 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 68% 993 928 CLONE COL03536. 73% 1186 998 HJABS31 825607 1627 WUblastx.64 (AAG49400) Ring-H2 protein. AAG49400 90% 1 1221 HJACE25 835517 1629 WUblastx.64 (O75921) RNA POLYMERASE II O75921 95% 381 662 TERMINATION FACTOR. 91% 64 390 HJACK21 664506 1630 WUblastx.64 (Q9CYB5) 5730552M22RIK Q9CYB5 98% 530 1123 PROTEIN. HJBCG74 839361 1631 WUblastx.64 (Q9UNW1) MULTIPLE INOSITOL Q9UNW1 88% 33 560 POLYPHOSPHATE 99% 560 1492 PHOSPHATASE. HJBCO21 695727 1632 WUblastx.64 (Q9Y6E5) HSPC024-ISO. Q9Y6E5 84% 9 482 HJBCQ40 831119 1633 WUblastx.64 (BAB55245) CDNA FLJ14722 fis, BAB55245 84% 58 1068 clone NT2RP3001621. HJBDM36 773168 1634 WUblastx.64 (O15498) SNARE PROTEIN YKT6. O15498 92% 127 720 HJMAZ60 844796 1637 WUblastx.64 (Q9D9R1) ADULT MALE TESTIS Q9D9R1 87% 482 1054 CDNA, RIKEN FULL-LENGTH 88% 14 484 ENRICHED LIBRARY, HJMBB20 824264 1638 WUblastx.64 ISOFORM GAC OF O94925 sp_vs|O94925- 99% 3 527 02|O94925 HJMBB20 842673 1639 WUblastx.64 ISOFORM GAC OF O94925 sp_vs|O94925- 100% 4 1461 02|O94925 HJMBB20 844890 1640 WUblastx.64 ISOFORM GAC OF O94925 sp_vs|O94925- 100% 4 1461 02|O94925 HJMBK59 664490 1641 WUblastx.64 (O75935) DYNACTIN SUBUNIT. O75935 100% 2 286 HJMBP01 638190 1642 WUblastx.64 (AAH00573) HSPC163 protein. AAH00573 100% 13 174 HJPAF69 589794 1646 WUblastx.64 (Q9H387) PRO2550. Q9H387 77% 439 531 73% 278 424 HJPAQ19 566832 1647 WUblastx.64 hypothetical protein pir|T08694|T08694 78% 9 869 DKFZp564O092.1 - human (fragment) HJPAZ35 845379 1648 WUblastx.64 (Q9Y262) HSPC025. Q9Y262 95% 126 1745 HJPBI77 602875 1649 WUblastx.64 (Q9V756) CG10155 PROTEIN. Q9V756 42% 16 249 HJPBN96 610087 1650 WUblastx.64 (Q9NX09) CDNA FLJ20500 FIS, Q9NX09 100% 106 288 CLONE KAT09159. 100% 70 108 HJPBU47 824063 1651 WUblastx.64 (Q9NPM0) 8D6 ANTIGEN Q9NPM0 100% 240 377 (FRAGMENT). 77% 315 380 73% 261 329 80% 329 844 HJPDK61 844034 1654 WUblastx.64 (Q9V7R7) CG8311 PROTEIN. Q9V7R7 41% 466 1194 HKABI53 805966 1655 WUblastx.64 (Q9BUP3) TAT-INTERACTING Q9BUP3 93% 126 212 PROTEIN (30 KD). 99% 212 778 HKABN63 566840 1656 WUblastx.64 (Q9BRZ8) SIMILAR TO RIKEN Q9BRZ8 85% 40 306 CDNA 1810009K13 GENE. 98% 263 700 HKACA25 824087 1657 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 64% 1493 1543 PROTEIN. 66% 1276 1488 HKACO64 897591 1658 WUblastx.64 (Q9P195) PRO1722. Q9P195 61% 694 593 58% 930 748 HKACX90 604042 1660 WUblastx.64 (Q9CZH5) 2700094L05RIK Q9CZH5 69% 13 510 PROTEIN. HKADI27 1028177 1661 WUblastx.64 (O35390) ENDO-ALPHA-D- O35390 65% 302 562 MANNOSIDASE. 51% 531 602 87% 574 1512 HKADN26 802131 1662 WUblastx.64 (Q24333) ELASTIN LIKE PROTEIN Q24333 48% 57 263 (FRAGMENT). HKADT55 668243 1664 WUblastx.64 (Q9CS80) 5730420G12RIK PROTEIN Q9CS80 97% 454 708 (FRAGMENT). 93% 13 57 HKAEK58 695729 1665 HMMER PFAM: Acyl-CoA dehydrogenase PF00441 449.8 160 1269 2.1.1 WUblastx.64 (Q9UKU7) ACYL-COENZYME A Q9UKU7 100% 39 113 DEHYDROGENASE-8. 99% 112 1284 HKAEK72 840457 1666 HMMER PFAM: Sugar (and other) transporter PF00083 70.8 270 575 2.1.1 WUblastx.64 (Q9UGQ3) SUGAR TRANSPORTER Q9UGQ3 99% 9 602 (GLUCOSE TRANSPORTER 9). HKAFQ41 847281 1668 WUblastx.64 (O94771) TRANSCRIPTION O94771 84% 36 896 FACTOR-LIKE 5. HKAHH71 846826 1669 WUblastx.64 (Q9VAA6) CG7950 PROTEIN. Q9VAA6 57% 31 348 HKAKU90 838608 1671 WUblastx.64 (Q9H2T6) ANTIGEN ART1/P17. Q9H2T6 85% 19 1230 HKFAA15 753275 1673 WUblastx.64 (Q9P195) PRO1722. Q9P195 67% 1748 1593 76% 1605 1465 HKGAJ81 581063 1676 WUblastx.64 (Q9H387) PRO2550. Q9H387 81% 445 335 48% 1240 1166 70% 620 429 HKGAK45 809093 1677 WUblastx.64 (Q9H387) PRO2550. Q9H387 63% 1946 1815 68% 2088 1945 HKGAP57 841627 1679 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 54% 770 639 CLONE KAIA0536. 72% 907 767 HKGAW41 858745 1680 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 68% 1458 1327 CLONE COL04765. 71% 1622 1431 HKGBA21 822870 1681 WUblastx.64 (O00367) L1 ELEMENT L1.19 P40 O00367 55% 1697 1783 PROTEIN. 30% 810 1064 50% 1279 1731 HKGBC33 823368 1682 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 76% 783 1001 HKGBC73 608169 1683 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 54% 74 9 PROTEIN. 78% 133 77 HKGBF61 658697 1684 WUblastx.64 (AAK55521) PRO0764. AAK55521 70% 978 919 39% 1700 1587 59% 138 73 70% 1171 980 HKGBH54 588482 1685 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 61% 2206 2114 43% 1994 1884 54% 2135 1959 HKGBP52 899282 1686 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 68% 1562 1440 CLONE KAIA0536. 76% 1726 1556 HKGCE23 564825 1687 WUblastx.64 (Q9D445) 4933414E04RIK Q9D445 60% 1456 1749 PROTEIN. HKGCE62 638200 1688 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 63% 995 897 61% 1131 985 HKGCK41 559327 1689 WUblastx.64 (Q9NQ89) MIB001 PROTEIN. Q9NQ89 93% 20 583 HKGCK41 795256 1690 WUblastx.64 (Q9NQ89) MIB001 PROTEIN. Q9NQ89 93% 31 591 HKGDA95 581064 1693 WUblastx.64 (Q9BYC7) MITOCHONDRIAL Q9BYC7 98% 77 313 RIBOSOMAL PROTEIN BMRP36A. 98% 618 782 82% 428 625 HKGDO12 857225 1694 WUblastx.64 (O60448) NEURONAL THREAD O60448 55% 1223 1170 PROTEIN AD7C-NTP. 72% 1429 1277 57% 1437 1222 48% 1218 1129 56% 1289 1146 79% 1441 1223 HKIME53 636982 1695 WUblastx.64 (Q9H3S2) HCCA2 PROTEIN. Q9H3S2 100% 4 30 100% 29 70 89% 63 446 HKIMG23 738671 1696 WUblastx.64 (O75896) FUS1 PROTEIN. FUS1_HUMAN 76% 217 255 100% 131 172 100% 246 458 HKMLF77 604743 1704 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 91% 673 638 CLONE KAIA0536. 88% 737 684 52% 627 457 HKMLM32 826523 1705 WUblastx.64 (Q9P0E3) HSPC093 (FRAGMENT). Q9P0E3 57% 861 1001 HKMLR17 847432 1706 HMMER PFAM: CUB domain PF00431 145 425 760 2.1.1 WUblastx.64 (Q9UKZ9) PROCOLLAGEN C- Q9UKZ9 94% 8 1210 TERMINAL PROTEINASE ENHANCER PROTEIN 2. HKMLT89 826167 1707 WUblastx.64 (Q9HD86) NAG18. Q9HD86 87% 1141 1233 HKMLV05 663924 1708 WUblastx.64 (Q9H387) PRO2550. Q9H387 81% 1559 1290 HKMLV25 695731 1709 WUblastx.64 (Q9BT42) SIMILAR TO LR8 Q9BT42 97% 328 474 PROTEIN. 40% 266 340 99% 2 334 HKMMD91 847394 1712 WUblastx.64 (AAH07729) Similar to RIKEN cDNA AAH07729 100% 1150 752 2810021O14 gene. HKMMP90 746876 1713 WUblastx.64 (O40947) ORF 73. O40947 30% 237 398 28% 240 443 30% 210 317 24% 1054 1569 25% 1054 1659 24% 1054 1659 25% 1054 1659 25% 1060 1662 23% 1054 1677 HKPAC10 527238 1715 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 47% 970 677 CLONE HEMBA1006036. HKPAC50 830367 1716 WUblastx.64 (O95139) NADH-UBIQUINONE NB7M_HUMAN 61% 106 324 OXIDOREDUCTASE B17 SUBUNIT (EC 1.6 HKTAC18 831292 1718 WUblastx.64 (Q9Y5W8) SORTING NEXIN 13 SNXD_HUMAN 88% 3 53 (FRAGMENT). 96% 38 400 HL1SA89 839594 1719 WUblastx.64 (Q9Y399) MITOCHONDRIAL 28S Q9Y399 99% 63 641 RIBOSOMAL PROTEIN S2 (MRP- S2). HL2AB60 722226 1720 WUblastx.64 SHORT ISOFORM OF Q9Y679 sp_vs|Q9Y679- 91% 64 1293 01|Q9Y679 HL3AF32 753468 1722 WUblastx.64 (Q9CZ73) 2810049G06RIK Q9CZ73 76% 6 527 PROTEIN. HLDAV70 623603 1723 WUblastx.64 (Q9UJW0) DYNACTIN P62 Q9UJW0 100% 175 285 SUBUNIT (CDNA FLJ20292 FIS, CLONE HEP05374). HLDBL62 899498 1724 WUblastx.64 (Q9NYC7) HEPATOCELLULAR Q9NYC7 78% 36 740 CARCINOMA-ASSOCIATED ANTIGEN 112. HLDBV18 805684 1725 WUblastx.64 (Q9H7D5) CDNA: FLJ21022 FIS, Q9H7D5 94% 284 1198 CLONE CAE06383. HLDBV54 570813 1726 HMMER PFAM: Sec1 family PF00995 18.7 180 332 2.1.1 HLDNF18 821599 1730 WUblastx.64 (Q9NWI8) CDNA FLJ20823 FIS, Q9NWI8 78% 17 583 CLONE ADSE00282 (SIMILAR TO TRANSLOCASE O HLDNN84 612801 1731 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 51% 751 849 59% 600 755 HLDOD77 610208 1732 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 71% 513 617 PROTEIN. 62% 395 523 HLDOL74 812765 1733 WUblastx.64 (Q9P0A2) HSPC279 (FRAGMENT). Q9P0A2 94% 57 671 HLDRU08 701995 1735 WUblastx.64 (Q9NVD7) CDNA FLJ10793 FIS, Q9NVD7 99% 13 729 CLONE NT2RP4000588 (ALPHA- PARVIN). HLDXF43 514022 1736 WUblastx.64 cytochrome-c oxidase (EC 1.9.3.1) pir|A00472|OBHU2 52% 164 559 chain II - human mitochondrion HLEAA10 824265 1737 WUblastx.64 (Q13579) MARINER Q13579 85% 842 741 TRANSPOSASE. 77% 738 13 HLHAE14 865625 1739 WUblastx.64 (Q9H7S6) CDNA FLJ14310 FIS, Q9H7S6 66% 1412 1528 CLONE PLACE3000271. 44% 935 1060 HLHAE14 886763 1740 WUblastx.64 (Q9H7S6) CDNA FLJ14310 FIS, Q9H7S6 66% 1412 1528 CLONE PLACE3000271. 44% 935 1060 HLHBS54 837503 1741 WUblastx.64 (Q9NZV5) SELENOPROTEIN N SELN_HUMAN 92% 1 1455 PRECURSOR. HLHCG24 847398 1744 WUblastx.64 (Q9BZE5) PGC-1 RELATED CO- Q9BZE5 82% 10 615 ACTIVATOR. HLHCN51 836150 1746 WUblastx.64 (Q9D1H8) 1110007K17RIK Q9D1H8 72% 1588 1514 PROTEIN. 48% 1514 1275 HLHCT96 886178 1747 WUblastx.64 (Q9H6G8) CDNA: FLJ22294 FIS, Q9H6G8 90% 1976 1917 CLONE HRC04426. 68% 2152 1955 HLHDJ05 841732 1750 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 68% 812 708 PROTEIN. 66% 708 547 HLHDL37 847399 1751 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 78% 690 571 CLONE KAT08285. HLHDL69 883335 1752 WUblastx.64 (O09014) PEPTIDE/HISTIDINE O09014 88% 305 592 TRANSPORTER. 72% 598 1314 HLHDL69 850691 1753 WUblastx.64 (O09014) PEPTIDE/HISTIDINE O09014 75% 305 340 TRANSPORTER. 78% 330 1484 HLHDL69 843836 1754 WUblastx.64 (O09014) PEPTIDE/HISTIDINE O09014 75% 305 340 TRANSPORTER. 78% 330 1484 HLHDL69 855906 1755 WUblastx.64 (O09014) PEPTIDE/HISTIDINE O09014 75% 305 340 TRANSPORTER. 78% 330 1484 HLHDM38 566843 1756 WUblastx.64 hypothetical protein pir|T46471|T46471 45% 6 326 DKFZp434L0130.1 - human 39% 181 339 61% 476 886 HLHDR92 692150 1757 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 77% 1252 1172 CLONE KAIA0536. 58% 1181 1095 HLHDY94 847121 1758 WUblastx.64 (O00572) HYPOTHETICAL 43.0 KDA O00572 56% 637 876 PROTEIN. 61% 18 752 HLHEI72 874180 1761 HMMER PFAM: Eukaryotic protein kinase PF00069 106.7 101 403 2.1.1 domain WUblastx.64 (Q9Y6S4) SIMILARITY IS TO Q9Y6S4 81% 589 621 SERINE/THREONINE-PROTEIN 97% 89 526 KINASE (FRAGMENT). HLHEX62 608285 1762 WUblastx.64 (Q9H387) PRO2550. Q9H387 70% 1102 833 HLHFP09 805967 1764 WUblastx.64 ISOFORM 2 OF O95460 sp_vs|O95460- 98% 3 881 01|O95460 38% 102 740 HLHGG78 638210 1765 WUblastx.64 (O95989) DIPHOSPHOINOSITOL O95989 100% 10 291 POLYPHOSPHATE PHOSPHOHYDROLASE. HLHSQ35 570814 1767 WUblastx.64 (Q9JIX6) RNA BINDING PROTEIN Q9JIX6 100% 671 712 NAPOR-3 (FRAGMENT). 87% 711 827 HLHTP55 847322 1769 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 50% 320 433 CLONE COL04765. 56% 462 635 HLIBD74 604914 1770 WUblastx.64 (Q9H797) CDNA: FLJ21128 FIS, Q9H797 98% 12 290 CLONE CAS06258. HLIBE41 772762 1771 WUblastx.64 (Q9D0Q3) 1300007B24RIK Q9D0Q3 58% 71 409 PROTEIN. HLLAX64 827292 1775 WUblastx.64 (BAB55004) CDNA FLJ14357 fis, BAB55004 96% 1020 1502 clone HEMBA1000005, h 91% 27 416 77% 364 417 25% 996 1196 97% 395 1057 32% 761 862 28% 752 973 28% 1257 1478 45% 611 676 45% 1428 1499 17% 1002 1229 HLLAX95 588405 1776 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 52% 541 603 PROTEIN. 65% 381 521 HLMCT51 772579 1780 WUblastx.64 (Q9DAD1) 1700013E09RIK Q9DAD1 55% 443 883 PROTEIN. HLMCT95 570815 1781 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 53% 1414 1121 PRODUCT. HLMDD65 566891 1782 WUblastx.64 (P20591) INTERFERON- MX1_HUMAN 100% 489 575 REGULATED RESISTANCE GTP- 70% 580 720 BINDING PROTEI HLMFB62 604915 1785 WUblastx.64 (Q9H700) CDNA: FLJ21617 FIS, Q9H700 68% 154 2 CLONE COL07481. HLMFG52 602166 1786 WUblastx.64 (Q9UHT5) PRO1848. Q9UHT5 71% 837 941 HLMFU53 801936 1787 WUblastx.64 (Q9ESD6) LNV. Q9ESD6 67% 1141 620 HLMHG68 580850 1788 WUblastx.64 (Q9HAI5) CDNA FLJ11583 FIS, Q9HAI5 92% 305 775 CLONE HEMBA1003680, WEAKLY SIMILAR TO PUT HLMHN06 853606 1789 WUblastx.64 probable transposase - human pir|S72481|S72481 59% 456 286 transposon MER37 80% 262 107 HLMIM84 588487 1791 WUblastx.64 (Q9Y3B0) CGI-105 PROTEIN. Q9Y3B0 84% 296 334 98% 645 806 HLMIW76 570816 1794 WUblastx.64 (O60448) NEURONAL THREAD O60448 85% 531 490 PROTEIN AD7C-NTP. 57% 528 472 59% 600 535 57% 528 472 77% 744 718 63% 601 536 66% 752 591 69% 736 590 70% 737 474 HLQAD72 853613 1798 HMMER PFAM: Pancreatic ribonucleases PF00074 213.4 208 564 2.1.1 WUblastx.64 ribonuclease 4 (EC 3.1.—.—) precursor - pir|I52489|I52489 90% 124 564 human HLQAM30 609880 1799 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 76% 648 610 57% 490 398 73% 617 480 HLQAM59 560662 1800 WUblastx.64 (AAH08373) Similar to hypothetical AAH08373 82% 879 829 protein PRO1722. 54% 754 722 63% 1018 881 HLQBB23 745364 1801 WUblastx.64 (Q9CYS6) 2810459M11RIK Q9CYS6 44% 38 286 PROTEIN. HLQCY09 745384 1804 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 57% 884 843 CLONE KAT08285. 58% 759 505 HLQDK45 621414 1807 WUblastx.64 (Q9CPU5) 2700019M19RIK Q9CPU5 100% 615 737 PROTEIN. HLQDM47 743237 1808 WUblastx.64 giant protein p619 - human pir|S71752|S71752 92% 377 616 HLTDI20 610269 1817 WUblastx.64 (Q9H6C9) CDNA: FLJ22385 FIS, Q9H6C9 70% 5 76 CLONE HRC07610. 95% 58 186 HLTDI65 740756 1818 WUblastx.64 (O60448) NEURONAL THREAD O60448 41% 1290 1382 PROTEIN AD7C-NTP. 50% 1456 1623 46% 1881 2027 47% 1881 2018 54% 1958 2029 54% 1878 1949 41% 1513 1605 57% 2095 2157 55% 1955 2038 60% 1275 1484 54% 1372 1464 47% 2095 2163 42% 2008 2157 64% 1472 1513 61% 1263 1463 36% 1939 2079 32% 1296 1460 50% 1363 1470 61% 2015 2149 46% 1315 1608 HLTDK30 834810 1819 WUblastx.64 (Q9HA72) CDNA FLJ12133 FIS, Q9HA72 100% 511 747 CLONE MAMMA1000278 97% 231 506 (SIMILAR TO HYPOTHETIC 97% 740 1180 HLTDL37 638213 1820 WUblastx.64 (Q9Y383) CGI-74 PROTEIN. Q9Y383 82% 2 754 66% 1084 1155 37% 974 1078 HLTDU35 827293 1821 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 44% 1118 954 PRODUCT. 62% 954 820 HLTEH84 782094 1823 WUblastx.64 (Q9H5P9) CDNA: FLJ23188 FIS, Q9H5P9 97% 15 491 CLONE LNG12038. HLTEL39 853615 1824 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 39% 1391 1218 CLONE COL03536. 70% 1536 1387 HLTEW52 663148 1826 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 70% 1498 1244 CLONE KAIA0536. HLTEZ36 821676 1827 WUblastx.64 (Q9H387) PRO2550. Q9H387 76% 7 171 HLTGG14 824068 1828 WUblastx.64 (Q9N032) UNNAMED PROTEIN Q9N032 70% 1142 1020 PRODUCT. HLUAF94 794116 1829 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 75% 1438 1473 72% 1256 1444 HLWAH33 855954 1830 WUblastx.64 (Q9NSI3) PRED43 PROTEIN Q9NSI3 100% 1660 1761 (FRAGMENT). 96% 25 102 HLWAO11 885339 1831 WUblastx.64 (Q9ES77) POLYDOM PROTEIN Q9ES77 76% 14 2656 PRECURSOR. 32% 14 2371 32% 2 2383 29% 152 2635 28% 14 2371 27% 14 1699 28% 506 1408 30% 2 628 29% 38 640 27% 161 787 28% 1469 2020 31% 2084 2617 34% 2267 2659 33% 14 373 32% 80 448 32% 2021 2635 35% 1019 1297 29% 1472 1969 22% 494 1990 33% 1745 2074 25% 14 478 27% 1067 1597 28% 1886 2179 26% 170 895 24% 1310 1771 26% 2249 2638 30% 1430 1654 22% 1142 1609 35% 1145 1288 36% 2003 2110 18% 1382 2194 35% 95 187 24% 455 709 27% 2354 2560 25% 440 673 33% 173 271 HLWAX50 809094 1833 WUblastx.64 (AAC08702) Meltrin-L precursor. AAC08702 92% 628 1629 93% 1587 2273 64% 178 255 39% 1553 1669 97% 257 661 HLWBK16 638217 1835 WUblastx.64 (Q9UEV9) ACTIN-BINDING Q9UEV9 94% 679 1191 PROTEIN HOMOLOG ABP-278. 28% 572 700 52% 590 658 57% 605 658 58% 593 658 34% 673 750 29% 682 1185 50% 593 652 37% 593 721 33% 727 1164 37% 853 1125 30% 664 1179 53% 590 775 31% 593 775 34% 730 1188 34% 691 1185 32% 745 1125 32% 590 775 37% 706 1185 32% 691 1170 32% 688 1179 37% 688 1173 37% 703 1185 31% 703 1182 28% 703 1176 36% 715 1149 30% 703 1188 32% 949 1173 32% 691 1158 28% 691 1185 41% 706 1185 36% 593 775 32% 593 775 26% 551 775 31% 691 1179 34% 593 775 30% 748 1125 HLYAJ79 608289 1839 WUblastx.64 (Q9B2U5) ATP SYNTHASE 6. Q9B2U5 63% 301 807 HLYAT54 581067 1842 WUblastx.64 (Q9H9F0) CDNA FLJ12802 FIS, Q9H9F0 93% 35 220 CLONE NT2RP2002124, WEAKLY SIMILAR TO UBI HLYBC81 588490 1843 WUblastx.64 (Q9HBW6) NAG13. Q9HBW6 43% 1373 1711 HLYBD09 610019 1844 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 69% 1546 1644 CLONE KAIA0536. 65% 1391 1513 HLYBL67 834538 1845 WUblastx.64 (Q9NRE8) PADI-H PROTEIN. Q9NRE8 64% 335 294 61% 304 158 HLYBN23 667995 1847 WUblastx.64 (Q9UI59) PRO0478 PROTEIN. Q9UI59 83% 1397 1450 80% 1333 1395 HLYBT28 608152 1850 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 54% 1082 774 PRODUCT. HLYBY04 596819 1852 WUblastx.64 (Q9H387) PRO2550. Q9H387 83% 1565 1530 71% 1533 1270 HLYCE15 621407 1853 WUblastx.64 (Q9P147) PRO2822. Q9P147 75% 518 420 HLYCH04 608295 1854 WUblastx.64 (O62658) LINE-1 ELEMENT ORF2. O62658 47% 106 273 45% 7 39 HLYDE38 735987 1856 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 47% 987 835 PRODUCT. 68% 1091 1005 HLYDG55 835625 1857 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 73% 638 423 CLONE LNG09295. HLYEA60 685469 1859 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 50% 104 181 70% 27 116 HLYEJ14 853362 1860 WUblastx.64 (O95498) VASCULAR NON- VNN2_HUMAN 97% 94 1653 INFLAMMATORY MOLECULE 2 PRECURSOR (VA HLYEJ44 838149 1861 WUblastx.64 (Q9H400) DJ583P15.4.1 (NOVEL Q9H400 100% 124 153 PROTEIN (TRANSLATION OF 80% 273 1007 CDNA FLJ20406 (E HLYEU51 607815 1862 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 67% 691 563 CLONE KAIA0536. 78% 857 708 HLYGV19 826334 1863 WUblastx.64 (Q9H0W4) HYPOTHETICAL 19.2 KDA Q9H0W4 72% 195 644 PROTEIN. HMABK52 853363 1864 WUblastx.64 (Q9N032) UNNAMED PROTEIN Q9N032 62% 1136 1002 PRODUCT. HMACL77 853365 1866 WUblastx.64 (O95564) HYPOTHETICAL 30.9 KDA O95564 100% 8 256 PROTEIN. HMACT74 843744 1867 WUblastx.64 gamma-interferon-inducible protein pir|A43708|A43708 94% 403 639 IP-30 precursor - human HMADJ14 843725 1868 WUblastx.64 (Q9H295) DC-SPECIFIC Q9H295 96% 871 945 TRANSMEMBRANE PROTEIN. 90% 47 880 HMADJ74 795479 1869 WUblastx.64 (Q9H295) DC-SPECIFIC Q9H295 100% 1010 1393 TRANSMEMBRANE PROTEIN. 93% 186 1067 HMAEA58 872563 1870 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 36% 584 519 CLONE KAIA0536. 59% 1339 1172 HMAGF01 837235 1871 WUblastx.64 (Q9UJ41) RAB5 GDP/GTP Q9UJ41 89% 71 1543 EXCHANGE FACTOR HOMOLOGUE. HMCED78 829071 1873 WUblastx.64 (Q9NWY5) CDNA FLJ20533 FIS, Q9NWY5 98% 380 862 CLONE KAT10931. HMCFN86 886182 1874 WUblastx.64 (Q9BVB2) UNKNOWN (PROTEIN Q9BVB2 100% 22 294 FOR IMAGE: 3459631) (FRAGMENT). HMCGJ47 699850 1875 WUblastx.64 (AAK29328) Serine AAK29328 91% 11 511 palmitoyltransferase. 91% 495 1430 HMCGK88 654854 1876 WUblastx.64 (Q9BT48) SIMILAR TO Q9BT48 100% 506 330 HYPOTHETICAL PROTEIN 92% 982 506 FLJ20116. HMCIH27 862038 1877 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 70% 1293 1063 CLONE COL04765. HMDAE88 638222 1881 WUblastx.64 (Q9H3C0) PRO0898. Q9H3C0 57% 1001 843 81% 844 734 HMDAM39 602907 1885 WUblastx.64 (Q9P195) PRO1722. Q9P195 66% 648 388 HMEAA41 587304 1886 WUblastx.64 (Q9H6H3) CDNA: FLJ22282 FIS, Q9H6H3 100% 9 419 CLONE HRC03861. HMEEH21 603187 1888 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 65% 280 504 PRODUCT. HMEEZ07 638225 1890 WUblastx.64 (Q9UK97) F-BOX ONLY PROTEIN FBX9_HUMAN 100% 795 917 9. HMEIH57 840370 1892 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 74% 238 11 PROTEIN (FRAGMENT). 58% 1348 1190 50% 1236 256 HMEIJ21 767276 1893 WUblastx.64 (Q9H067) HYPOTHETICAL 66.5 KDA Q9H067 61% 2388 1777 PROTEIN. 50% 828 709 56% 1676 840 HMEJC96 761218 1895 WUblastx.64 (O60836) MEMBRANE O60836 86% 193 669 GLYCOPROTEIN GP36 PRECURSOR. HMEJK28 825591 1897 WUblastx.64 (O00479) NON-HISTONE O00479 54% 273 485 CHROMOSOMAL PROTEIN (HIGH- MOBILITY GROUP (NONHIS HMEKW44 825499 1899 WUblastx.64 (O18975) HYPOTHETICAL 16.6 KDA O18975 71% 7 198 PROTEIN (FRAGMENT). HMEKW71 855955 1900 WUblastx.64 (Q9UHF1) NOTCH4-LIKE PROTEIN Q9UHF1 96% 579 758 (HYPOTHETICAL 29.6 KDA PROTEIN). HMHBI09 847402 1903 WUblastx.64 (AAH07644) Similar to RIKEN cDNA AAH07644 93% 39 683 9430029K10 gene (F HMHBI93 844729 1904 WUblastx.64 (Q9Z1S4) TGFB1-INDUCED ANTI- TIAF_MOUSE 90% 1391 1735 APOPTOTIC FACTOR 1 (12 KDA TGF- HMIAC52 825484 1906 WUblastx.64 (AAH00713) Neighbor of A-kinase AAH00713 30% 941 1201 anchoring protein 9 52% 988 1056 75% 112 927 HMIAG42 846339 1908 WUblastx.64 (AAK54122) RGS20 ret splice variant AAK54122 78% 1071 1730 1. HMIAG55 736020 1909 WUblastx.64 (AAD20460) Ribosomal protein L11. AAD20460 67% 424 609 HMIAG72 824091 1910 WUblastx.64 (O60519) CRE BINDING PROTEIN- O60519 85% 318 677 LIKE 2. HMIAO82 833070 1912 WUblastx.64 hypothetical protein pir|T46340|T46340 58% 32 358 DKFZp434B0814.1 - human 50% 443 742 (fragment) HMIAR42 809096 1913 WUblastx.64 (Q9BVE4) SIMILAR TO Q9BVE4 87% 1046 1165 INTERFERON-RELATED 91% 831 1052 DEVELOPMENTAL REGULATOR 99% 7 849 1. HMIAV33 668252 1914 WUblastx.64 (Q9UN80) HYPOTHETICAL 149.0 KDA Q9UN80 34% 1051 1182 PROTEIN. 48% 1167 1337 40% 603 430 41% 138 10 28% 1015 611 38% 431 162 HMIBE95 804597 1917 WUblastx.64 probable transposase - human pir|S72481|S72481 60% 1932 2075 transposon MER37 64% 1221 1355 42% 1355 1948 HMKAN71 884161 1920 WUblastx.64 (Q96G03) Unknown (protein for Q96G03 100% 12 1385 MGC: 19508). HMKBA33 596949 1921 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 47% 1147 1010 PRODUCT. 44% 1248 1162 75% 1441 1382 57% 1314 1147 33% 383 261 65% 1691 1482 HMKCK32 561576 1923 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 53% 619 335 PRODUCT. HMKDG69 588407 1927 WUblastx.64 pro-pol-dUTPase polyprotein - murine pir|T29097|T29097 59% 104 24 endogenous retrovirus ERV-L 34% 700 116 (fragment) HMKDM80 608168 1928 WUblastx.64 (Q9H6G8) CDNA: FLJ22294 FIS, Q9H6G8 74% 178 11 CLONE HRC04426. HMKEG88 634245 1929 WUblastx.64 (O00378) PUTATIVE P150. O00378 47% 294 4 HMMAA09 822859 1930 WUblastx.64 (AAK55521) PRO0764. AAK55521 68% 1305 1249 67% 1271 1014 HMMAK92 610100 1931 WUblastx.64 (Q9CZ33) 2810408E11RIK Q9CZ33 86% 15 152 PROTEIN. HMMBF22 780406 1934 WUblastx.64 (Q9H6V7) CDNA: FLJ21825 FIS, Q9H6V7 56% 1228 1341 CLONE HEP01348. 93% 407 499 58% 824 1159 HMMBH94 607463 1936 WUblastx.64 (AAK55521) PRO0764. AAK55521 80% 857 813 71% 835 593 HMMBK55 608179 1937 WUblastx.64 (Q9H397) PRO2852. Q9H397 72% 521 468 36% 480 361 70% 691 527 HMMBT47 804594 1941 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 66% 1238 1203 CLONE KAIA0536. 54% 1206 874 HMMCD35 607840 1942 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 59% 813 676 CLONE COL04765. 58% 661 512 HMMCD95 603202 1943 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 58% 1498 1397 PRODUCT. 63% 1643 1494 HMPAB26 580854 1944 WUblastx.64 (P78560) DEATH DOMAIN CRAD_HUMAN 100% 66 365 CONTAINING PROTEIN CRADD (CASPASE AND HMQAI38 589964 1946 WUblastx.64 immune-responsive gene 1 - mouse pir|I54546|I54546 71% 11 1363 (fragment) HMQAT69 799551 1947 WUblastx.64 (Q9Y5X1) SORTING NEXIN 9 (SH3 SNX9_HUMAN 94% 739 1842 AND PX DOMAIN-CONTAINING 82% 660 809 PROT 93% 56 748 HMQBL90 600363 1948 WUblastx.64 pre-B cell Ig lambda-like omega light pir|A33911|A33911 52% 879 1091 chain (non-rearranging) 14.1 - human 76% 974 1309 HMQBV82 826522 1949 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 61% 739 876 PROTEIN. 72% 969 868 40% 651 592 HMQCA75 767086 1950 WUblastx.64 (AAK55521) PRO0764. AAK55521 62% 1235 1071 72% 1072 944 HMQCB37 732168 1951 WUblastx.64 (AAK55521) PRO0764. AAK55521 85% 426 446 71% 1087 1149 50% 872 1042 79% 503 904 HMQCX41 603388 1953 WUblastx.64 (O60448) NEURONAL THREAD O60448 43% 192 260 PROTEIN AD7C-NTP. 55% 16 204 HMQDM09 664494 1954 WUblastx.64 (Q9NZV6) SELENOPROTEIN X 1 SELX_HUMAN 98% 38 385 (PROTEIN HSPC270). HMSAP33 731969 1956 WUblastx.64 (Q9P147) PRO2822. Q9P147 83% 1526 1398 HMSAZ48 877650 1957 WUblastx.64 probable pol polyprotein-related pir|S21348|S21348 46% 793 1218 protein 4 - rat 66% 722 784 HMSBN18 847405 1958 WUblastx.64 (O95789) ZNF258. O95789 65% 165 329 HMSBS25 826487 1959 WUblastx.64 (Q9Y642) K: CL COTRANSPORTER Q9Y642 77% 1019 1189 3. 81% 337 384 100% 257 346 HMSBU14 844443 1960 WUblastx.64 hypothetical protein pir|T47135|T47135 41% 954 1184 DKFZp761L0812.1 - human 63% 1192 1488 (fragment) HMSCB94 847406 1962 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 45% 1690 1589 CLONE KAT08285. 80% 1931 1683 HMSCK12 825475 1963 WUblastx.64 (Q9P195) PRO1722. Q9P195 62% 771 613 81% 622 491 HMSCV75 838530 1965 WUblastx.64 (Q9NRE8) PADI-H PROTEIN. Q9NRE8 40% 1565 1461 67% 1438 1301 HMSCW44 716249 1967 WUblastx.64 (Q9P195) PRO1722. Q9P195 47% 1251 1102 66% 1112 942 HMSCZ19 847408 1968 WUblastx.64 (O60427) BC269730_2 O60427 84% 621 1124 (HYPOTHETICAL 52.0 KDA 94% 1 675 PROTEIN) (FATTY ACID DESAT HMSDI67 827298 1969 WUblastx.64 probable pol polyprotein-related pir|S21348|S21348 55% 1255 1121 protein 4-rat 38% 1748 1284 HMSDR28 801948 1971 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 76% 1097 1035 CLONE COL04765. 72% 1317 1102 HMSFT25 581069 1972 WUblastx.64 (AAK55521) PRO0764. AAK55521 43% 1088 1198 61% 918 1025 HMSFW52 603177 1973 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 67% 1380 1631 CLONE KAIA0536. HMSGU30 847409 1975 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 55% 1718 1494 PROTEIN. HMSHB42 792385 1976 WUblastx.64 (Q9NR81) RHOGEF Q9NR81 97% 4 867 (HYPOTHETICAL 59.8 KDA PROTEIN). HMSHB42 600401 1977 WUblastx.64 (Q9NR81) RHOGEF Q9NR81 99% 4 1203 (HYPOTHETICAL 59.8 KDA PROTEIN). HMSHN72 855917 1978 WUblastx.64 (Q98SS5) DOUBLECORTIN. Q98SS5 91% 650 754 HMSHW73 704099 1980 WUblastx.64 (Q9P195) PRO1722. Q9P195 62% 2214 2134 65% 2131 2063 71% 2072 1935 HMSII36 825453 1982 WUblastx.64 (AAK55521) PRO0764. AAK55521 61% 23 61 72% 51 161 HMSIT42 581070 1983 WUblastx.64 (Q9P147) PRO2822. Q9P147 80% 1160 936 HMSJB08 877651 1984 WUblastx.64 (Q9D2Y4) 9130019I15RIK Q9D2Y4 56% 1037 612 PROTEIN. 58% 1987 1028 HMSJR44 638098 1987 WUblastx.64 (Q9P195) PRO1722. Q9P195 73% 798 721 43% 1168 1037 82% 853 803 36% 1633 1511 64% 995 843 HMSKQ91 825450 1988 WUblastx.64 (Q9HBN2) HYPOTHETICAL 15.8 KDA Q9HBN2 68% 864 742 PROTEIN. HMTAF92 740784 1990 WUblastx.64 (Q9Y6I4) UBIQUITIN CARBOXYL- UBP3_HUMAN 93% 2211 2167 TERMINAL HYDROLASE 3 (EC 84% 2053 644 3.1.2. HMTAT36 811885 1991 WUblastx.64 (Q9H6Z9) CDNA: FLJ21620 FIS, Q9H6Z9 100% 1 117 CLONE COL07838. HMUAD65 809097 1993 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 67% 1524 1213 CLONE COL03536. HMUBK53 553624 1997 WUblastx.64 (Q9H8X7) CDNA FLJ13156 FIS, Q9H8X7 86% 18 398 CLONE NT2RP3003490, WEAKLY SIMILAR TO HOM HMUBY57 844800 2001 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 78% 1827 1609 CLONE LNG09295. HMUBZ15 609837 2002 WUblastx.64 (Q9BU61) SIMILAR TO NUCLEAR Q9BU61 60% 968 1012 PROTEIN E3-3 ORF1. 100% 412 963 HMVAL15 822860 2003 WUblastx.64 A-kinase anchor protein 95 - human pir|T13161|T13161 83% 1 348 HMVBC84 743191 2004 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 47% 1019 957 CLONE COL04765. 76% 1135 1019 HMVBD68 831130 2005 WUblastx.64 catalase (EC 1.11.1.6) - pir|I40767|I40767 85% 207 103 Campylobacter jejuni HMVCG17 846161 2006 WUblastx.64 (Q9NZS9) APOPTOSIS Q9NZS9 96% 282 458 REGULATOR. 33% 89 151 100% 204 290 HMVDT89 831127 2011 WUblastx.64 (Q9HAT2) SIALIC ACID-SPECIFIC Q9HAT2 90% 2 385 ACETYLESTERASE II. 98% 384 809 HMVDT89 875313 2012 WUblastx.64 (Q9HAT2) SIALIC ACID-SPECIFIC Q9HAT2 90% 2 385 ACETYLESTERASE II. 98% 384 809 HMWAO65 839268 2013 WUblastx.64 (Q9P0A1) HSPC280 (FRAGMENT). Q9P0A1 100% 30 371 HMWAO82 825427 2014 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 84% 461 366 CLONE KAT08285. HMWBK35 566888 2016 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 76% 956 918 CLONE KAT08285. 64% 919 650 HMWBL38 602940 2018 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 76% 743 615 CLONE KAT08285. 65% 916 719 HMWBM48 566828 2019 WUblastx.64 ribosomal protein L3 precursor, pir|A27294|R5HUL3 80% 609 379 mitochondrial - human 100% 794 609 HMWCG28 847413 2020 WUblastx.64 (Q9P1S9) KINASE DEFICIENT Q9P1S9 84% 35 892 PROTEIN KDP (FRAGMENT). HMWCP85 757974 2021 WUblastx.64 Na+-dependent vitamin C (L-ascorbic pir|JC7182|JC7182 64% 37 297 acid) transporter SVCT1 - human 79% 1181 1507 61% 698 1264 86% 369 737 HMWDG30 623758 2022 WUblastx.64 (Q9VQ76) CG4263 PROTEIN. Q9VQ76 43% 1364 1411 37% 30 668 HMWDU20 695743 2023 WUblastx.64 (Q9NXB8) CDNA FLJ20335 FIS, Q9NXB8 89% 918 1034 CLONE HEP11429 (FRAGMENT). 75% 347 454 93% 1004 1819 HMWDZ63 608146 2025 WUblastx.64 (P41565) ISOCITRATE IDHG_RAT 33% 399 301 DEHYDROGENASE [NAD] 100% 169 74 SUBUNIT GAMMA, MITO HMWEA77 839221 2026 WUblastx.64 (Q9BGX7) HYPOTHETICAL 13.0 KDA Q9BGX7 60% 1066 1110 PROTEIN. 70% 1100 1282 HMWEC03 598902 2027 WUblastx.64 (Q9P0X2) HEAT SHOCK PROTEIN Q9P0X2 100% 1066 1365 HSP60. 91% 573 1064 99% 105 572 HMWEF46 825433 2028 WUblastx.64 (Q9H0G8) HYPOTHETICAL 14.0 KDA Q9H0G8 100% 1113 1226 PROTEIN. HMWEK43 847414 2029 WUblastx.64 (Q9H8K0) CDNA FLJ13520 FIS, Q9H8K0 46% 940 1167 CLONE PLACE1005828. HMWEM23 863105 2030 WUblastx.64 (Q96QF0) SSX2 interacting protein Q96QF0 96% 1086 1532 hRabin3A, isoform alpha2. 96% 128 1114 HMWEM23 859387 2031 WUblastx.64 (Q9H673) CDNA: FLJ22548 FIS, Q9H673 95% 1086 1532 CLONE HSI00519. 96% 152 1114 HMWEU96 1310888 2033 WUblastx.64 (Q9UH62) HYPOTHETICAL 42.5 KDA Q9UH62 88% 91 1227 PROTEIN (ALEX3) (ALEX3 PROTEIN). HMWEU96 601698 3110 WUblastx.64 (AB039669) ALEX3 [Homo sapiens] dbj|BAA94602.1| 88% 86 1222 HMWEX02 596821 2034 WUblastx.64 (Q9H7H4) FLJ00116 PROTEIN Q9H7H4 67% 509 1000 (FRAGMENT). HMWFB65 638102 2035 WUblastx.64 (O95427) MCD4P HOMOLOG. O95427 74% 37 591 95% 894 959 100% 611 898 68% 9 59 HMWFD77 610105 2036 WUblastx.64 (Q9P1N7) PRO0974. Q9P1N7 83% 459 494 55% 293 433 HMWFO25 566855 2037 WUblastx.64 (Q9HA89) CDNA FLJ12060 FIS, Q9HA89 100% 742 816 CLONE HEMBB1002142. HMWGM41 847415 2039 WUblastx.64 (Q9H8P0) CDNA FLJ13352 FIS, Q9H8P0 84% 33 986 CLONE OVARC1002165, WEAKLY SIMILAR TO 3-O HMWGO95 582078 2040 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 82% 1446 1396 CLONE COL04765. 68% 1390 1145 HMWGV85 847416 2041 WUblastx.64 (Q9H8H3) CDNA FLJ13631 FIS, Q9H8H3 100% 41 298 CLONE PLACE1011090, HIGHLY SIMILAR TO HOM HMWGZ42 824259 2042 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 50% 262 309 FOR MGC: 5149). 42% 1316 1534 HMWIQ26 722234 2045 WUblastx.64 (Q9NGC3) CENTAURIN GAMMA Q9NGC3 48% 844 395 1A PROTEIN. 47% 298 5 43% 221 162 21% 388 218 HMWIU49 800894 2046 WUblastx.64 (O95831) PROGRAMED CELL PCD8_HUMAN 98% 3 407 DEATH PROTEIN 8, 38% 1384 1500 MITOCHONDRIAL PREC 94% 379 1362 HMWJJ62 838612 2047 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 71% 657 875 PROTEIN. HMWJJ64 581071 2048 WUblastx.64 (Q9D0Q7) 2600005P05RIK Q9D0Q7 78% 12 878 PROTEIN. HNAAD76 596801 2049 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 65% 1650 1528 PRODUCT. 59% 1807 1646 HNALD94 844722 2051 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 70% 764 817 CLONE KAIA0536. 72% 816 926 HNALE44 785045 2052 WUblastx.64 (Q9Y503) FILAMIN, MUSCLE Q9Y503 100% 12 86 ISOFORM. 30% 916 1170 72% 12 86 62% 12 83 52% 12 62 31% 783 896 60% 18 86 51% 12 95 45% 12 77 36% 18 92 36% 9 95 95% 79 1170 31% 916 1170 28% 49 795 36% 1096 1170 33% 1127 1207 35% 901 1170 31% 112 801 30% 1823 1927 29% 925 1170 39% 916 1170 30% 916 1170 27% 217 504 29% 18 98 46% 15 86 29% 910 1170 30% 901 1170 34% 79 792 42% 79 798 25% 940 1170 30% 79 792 32% 916 1170 38% 340 864 29% 88 855 30% 88 792 32% 109 795 33% 16 798 26% 103 855 29% 916 1170 25% 925 1170 33% 85 792 30% 262 795 29% 109 816 34% 916 1164 32% 520 792 26% 751 1170 29% 430 792 33% 940 1170 36% 12 77 31% 82 792 34% 79 783 HNEAK38 631279 2056 WUblastx.64 (Q26195) PVA1 GENE. Q26195 71% 1204 1100 47% 1383 1333 59% 1358 1197 HNEAK65 604991 2057 WUblastx.64 (Q9P195) PRO1722. Q9P195 72% 570 427 82% 715 560 HNEBY79 841769 2059 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 58% 944 1153 72% 1270 1488 HNECL75 782493 2061 WUblastx.64 (Q9HCX0) PELLINO. Q9HCX0 96% 215 649 100% 627 1268 HNECX90 834776 2062 WUblastx.64 (Q9UJL8) HYPOTHETICAL 43.5 KDA Q9UJL8 84% 81 1265 PROTEIN. HNECX90 881133 2063 WUblastx.64 (Q9UJL8) HYPOTHETICAL 43.5 KDA Q9UJL8 84% 81 1265 PROTEIN. HNEDP75 581072 2065 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 63% 910 722 CLONE KAIA0536. HNEDQ02 846061 2066 WUblastx.64 (Q9HA25) CDNA FLJ12374 FIS, Q9HA25 76% 941 1063 CLONE MAMMA1002470, 93% 1018 2013 WEAKLY SIMILAR TO PRO HNEDU46 695745 2067 WUblastx.64 (Q9H3P1) 6-PHOSPHOFRUCTO-2- Q9H3P1 100% 343 420 KINASE HEART ISOFORM. 95% 137 208 95% 766 825 HNFAD50 839369 2068 WUblastx.64 (Q9H175) HYPOTHETICAL 59.6 KDA Q9H175 47% 128 859 PROTEIN. HNFAD50 843242 2069 WUblastx.64 (Q9H175) HYPOTHETICAL 59.6 KDA Q9H175 54% 50 379 PROTEIN. HNFAG67 804546 2070 WUblastx.64 (O60759) CYTOHESIN BINDING O60759 73% 327 857 PROTEIN HE. HNFCJ77 886185 2071 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 74% 2342 2250 FOR MGC: 5149). 63% 1819 1763 73% 1763 1719 61% 2034 1792 HNFCY57 877653 2073 WUblastx.64 (AAL12497) Cryopyrin. AAL12497 91% 8 2203 HNFDL89 823362 2074 WUblastx.64 (Q9BT07) UNKNOWN (PROTEIN Q9BT07 46% 2098 2193 FOR MGC: 4033). 100% 1964 2101 HNFDU92 602933 2076 WUblastx.64 (O60448) NEURONAL THREAD O60448 60% 703 407 PROTEIN AD7C-NTP. 48% 721 425 88% 435 409 74% 570 388 56% 764 627 56% 522 388 43% 571 503 43% 495 388 57% 745 683 48% 757 677 HNFDY09 843300 2077 WUblastx.64 (Q9BZ41) BA476I15.3 (NOVEL Q9BZ41 92% 1243 1281 PROTEIN SIMILAR TO SEPTIN) 75% 1280 1363 (FRAGMENT). 53% 1104 1238 HNFDY31 724955 2078 WUblastx.64 (O00571) DEAD-BOX PROTEIN 3 DDX3_HUMAN 64% 169 621 (HELICASE-LIKE PROTEIN 2) 98% 693 1028 (HLP2 HNFEA17 753242 2079 WUblastx.64 (Q9BRG9) UNKNOWN (PROTEIN Q9BRG9 94% 965 1318 FOR MGC: 11314). HNFEP55 722236 2080 WUblastx.64 (Q9NW80) HYPOTHETICAL 73.6 KDA Q9NW80 41% 877 966 PROTEIN. 65% 158 862 HNFET12 824170 2081 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 75% 1217 1182 CLONE COL03536. 75% 1182 952 HNFFR59 634649 2082 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 65% 1613 1485 CLONE MAMMA1000472. HNFGR15 844044 2084 WUblastx.64 (Q99770) HYPOTHETICAL 15.4 KDA Q99770 53% 1128 1172 PROTEIN. 90% 824 886 HNFGW53 589525 2086 WUblastx.64 (Q9NUM6) CDNA FLJ11267 FIS, Q9NUM6 53% 777 938 CLONE PLACE1009174. HNFHV68 567445 2089 WUblastx.64 pro-pol-dUTPase polyprotein - murine pir|T29097|T29097 44% 380 24 endogenous retrovirus ERV-L (fragment) HNFIE15 898153 2090 WUblastx.64 (Q9H387) PRO2550. Q9H387 72% 2374 2300 62% 2576 2370 HNFIE29 852233 2091 WUblastx.64 (Q9HBS7) HYPOTHETICAL 14.2 KDA Q9HBS7 42% 1484 1407 PROTEIN. 54% 1419 1168 HNFJE27 825400 2093 WUblastx.64 (Q9NUQ9) CDNA FLJ11197 FIS, Q9NUQ9 93% 986 1123 CLONE PLACE1007690 (HYPOTHETICAL 36.7 KDA HNGAC71 695746 2095 WUblastx.64 (Q9GKW4) HYPOTHETICAL 12.6 KDA Q9GKW4 77% 64 11 PROTEIN. 85% 138 79 HNGAX06 841890 2099 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 100% 22 108 PROTEIN (FRAGMENT). HNGBE44 825391 2103 WUblastx.64 (O00437) AXONEMAL DYNEIN O00437 71% 323 502 HEAVY CHAIN (FRAGMENT). HNGBI83 566815 2105 WUblastx.64 (O60448) NEURONAL THREAD O60448 69% 322 420 PROTEIN AD7C-NTP. 53% 122 160 73% 331 420 74% 13 87 59% 10 87 51% 12 104 55% 22 102 61% 42 80 55% 28 81 HNGCF29 580201 2111 WUblastx.64 (Q9H387) PRO2550. Q9H387 71% 1112 1074 60% 1069 842 HNGCF64 823147 2112 WUblastx.64 (AAH07558) Unknown (protein for AAH07558 48% 1893 1765 MGC: 15483). 55% 1763 1629 HNGDF54 702002 2113 WUblastx.64 (Q9N8T2) POSSIBLE SRPA. Q9N8T2 29% 13 654 27% 14 679 27% 14 679 29% 13 654 HNGDH27 581074 2115 WUblastx.64 (Q92722) RETINOBLASTOMA 1. Q92722 45% 290 99 HNGDN07 847081 2116 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 80% 1091 966 HNGEE06 689506 2122 WUblastx.64 (Q9H960) CDNA FLJ12988 FIS, Q9H960 43% 66 1 CLONE NT2RP3000080. 60% 1040 909 HNGEN32 621308 2128 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 76% 361 399 CLONE LNG09295. 80% 408 557 HNGET33 825378 2131 WUblastx.64 (Q9H387) PRO2550. Q9H387 78% 459 755 HNGEZ02 825376 2134 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 81% 840 938 CLONE KAIA0536. HNGEZ90 634858 2135 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 72% 1119 1045 CLONE COL04765. 71% 956 810 HNGFD30 612782 2138 WUblastx.64 (O75423) O75423 72% 264 100 ORF3, SPLICEVARIANT_A. HNGFD31 663532 2139 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 76% 574 624 CLONE MAMMA1000472. 78% 491 574 HNGFG04 772548 2141 WUblastx.64 (Q9Y485) X-LIKE 1 PROTEIN. Q9Y485 100% 423 533 HNGFI21 581075 2145 WUblastx.64 (Q9EQC8) PAPILLARY RENAL Q9EQC8 58% 117 296 CELL CARCINOMA-ASSOCIATED PROTEIN. HNGFT70 581079 2150 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 76% 1073 855 CLONE LNG09295. HNGGF13 638117 2153 WUblastx.64 hypothetical protein hc1 - mouse pir|S26689|S26689 38% 13 183 (fragment) 37% 13 186 HNGGK63 822862 2154 WUblastx.64 (Q9QZ22) OLFACTORY Q9QZ22 59% 1224 1003 RECEPTOR. 88% 985 278 HNGGT10 866179 2159 HMMER PFAM: 3-beta hydroxysteroid PF01073 263.9 19 468 2.1.1 dehydrogenase/isomerase family WUblastx.64 (Q9BSN9) 3 BETA-HYDROXY- Q9BSN9 75% 10 480 DELTA 5-C27-STEROID OXIDOREDUCTASE. HNGHB89 722238 2161 WUblastx.64 (O88748) E-STOP PROTEIN. O88748 100% 153 37 HNGHD07 608149 2162 WUblastx.64 (Q9BRA2) UNKNOWN (PROTEIN Q9BRA2 51% 588 755 FOR MGC: 14353). 70% 746 943 HNGIK07 825360 2168 WUblastx.64 (Q9H3I6) BRAIN MY040 PROTEIN. Q9H3I6 76% 1139 912 HNGIM40 747699 2169 WUblastx.64 (Q9H5W7) CDNA: FLJ22921 FIS, Q9H5W7 87% 2354 2208 CLONE KAT06711. HNGIM83 751714 2170 HMMER PFAM: HCO3-transporter family PF00955 117.9 72 290 2.1.1 WUblastx.64 (Q9UIB9) BICARBONATE Q9UIB9 74% 9 290 TRANSPORTER. HNGIO93 897527 2171 HMMER PFAM: Ribosomal L27 protein PF01016 71.7 −21 −233 2.1.1 WUblastx.64 (Q9P0M9) HSPC250 Q9P0M9 88% 151 594 (MITOCHONDRIAL RIBOSOMAL PROTEIN L27) (L27MT) (HYPOT HNGIU16 826717 2173 WUblastx.64 (Q9H387) PRO2550. Q9H387 78% 2322 2215 76% 2505 2467 78% 2467 2315 HNGIX91 564574 2174 WUblastx.64 (AAH07609) Similar to hypothetical AAH07609 100% 956 982 protein PRO1722. 77% 885 950 76% 739 891 HNGJU60 825336 2182 WUblastx.64 (Q9GW02) EXTREMELY Q9GW02 51% 166 258 CYSTEINE/VALINE RICH 50% 166 261 PROTEIN (FRAGMENT). 46% 166 261 56% 169 258 48% 169 261 45% 169 261 48% 169 261 40% 290 487 44% 287 487 41% 290 487 41% 169 261 44% 287 490 46% 169 258 55% 231 257 44% 290 487 45% 287 487 42% 287 487 44% 290 487 44% 290 490 48% 163 261 46% 166 261 48% 169 261 48% 163 261 44% 287 487 40% 172 261 45% 169 261 43% 287 487 48% 169 261 HNGKW35 899408 2184 WUblastx.64 (O00627) HYPOTHETICAL 11.0 KDA O00627 53% 292 477 PROTEIN (ORF2). HNGKY94 835021 2185 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 66% 581 501 CLONE COL04765. 69% 777 583 HNHAB38 609892 2188 WUblastx.64 (Q9H387) PRO2550. Q9H387 68% 635 339 HNHAD34 612845 2190 WUblastx.64 (Q9N032) UNNAMED PROTEIN Q9N032 64% 1056 922 PRODUCT. HNHAY26 704789 2198 WUblastx.64 (Q9H397) PRO2852. Q9H397 85% 2098 1919 HNHAZ20 839696 2200 WUblastx.64 (Q9C074) HYPOTHETICAL 73.9 KDA Q9C074 81% 1213 11 PROTEIN (FRAGMENT). HNHBE38 562730 2202 WUblastx.64 (AAK55521) PRO0764. AAK55521 65% 1131 1072 72% 1097 855 HNHBG18 604924 2203 WUblastx.64 (O00365) L1 ELEMENT L1.15 P40 O00365 29% 717 271 PROTEIN. HNHBM16 566866 2205 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 87% 80 33 CLONE COL03536. 71% 310 80 HNHCH78 694657 2206 WUblastx.64 (AAH07609) Similar to hypothetical AAH07609 100% 270 290 protein PRO1722. 88% 194 271 75% 49 195 HNHCT47 634691 2210 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 46% 434 396 CLONE COL04765. 56% 621 448 HNHDC52 812424 2213 WUblastx.64 (O00365) L1 ELEMENT L1.15 P40 O00365 32% 1038 910 PROTEIN. 68% 904 269 HNHDI17 738043 2216 WUblastx.64 (O95662) POT. ORF VI O95662 80% 265 327 (FRAGMENT). 56% 5 295 HNHDR57 638121 2219 WUblastx.64 (O60448) NEURONAL THREAD O60448 58% 1193 1095 PROTEIN AD7C-NTP. 63% 496 440 56% 485 393 50% 503 426 64% 1195 1124 33% 658 440 41% 1186 1043 40% 402 322 61% 1065 874 51% 1177 923 36% 486 331 31% 1090 455 53% 437 393 60% 1168 863 HNHDW34 607867 2222 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 62% 1175 1017 CLONE COL04765. 56% 961 671 HNHEF70 610115 2228 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 80% 481 437 CLONE COL04765. 77% 417 352 50% 650 462 HNHEL22 607421 2231 WUblastx.64 (Q9NUL2) CDNA FLJ11292 FIS, Q9NUL2 71% 280 342 CLONE PLACE1009665. 72% 363 656 HNHEN70 854712 2232 WUblastx.64 olfactory receptor OR18 - rat pir|S29710|S29710 67% 34 567 HNHEP41 638124 2234 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 41% 915 730 CLONE KAIA0536. 73% 736 635 HNHEZ76 618544 2238 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 72% 359 466 CLONE MAMMA1000472. HNHFF81 732075 2240 WUblastx.64 (Q9GMP5) HYPOTHETICAL 6.6 KDA Q9GMP5 71% 1208 1333 PROTEIN. HNHFR42 638127 2242 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 75% 522 427 HNHGD95 609905 2244 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 46% 430 392 CLONE COL04765. 56% 608 444 HNHGR82 617117 2245 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 48% 125 39 PROTEIN. 56% 262 110 HNHGY77 638128 2247 WUblastx.64 (Q9BZV1) UBX DOMAIN- Q9BZV1 53% 292 248 CONTAINING PROTEIN 1. 100% 212 90 HNHHA47 658743 2248 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 65% 983 864 CLONE KAIA0536. 66% 1138 1001 HNHKI74 777856 2252 WUblastx.64 (Q9BGX7) HYPOTHETICAL 13.0 KDA Q9BGX7 64% 350 541 PROTEIN. HNHLD80 839255 2254 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 44% 563 414 CLONE MAMMA1000472. 72% 700 572 HNHLS76 853372 2255 WUblastx.64 (O15410) CAGH45. O15410 74% 146 316 HNHMY76 838256 2259 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 75% 576 647 CLONE KAIA0536. 83% 520 555 75% 659 778 HNKAA76 802009 2264 WUblastx.64 (Q9BTI4) SIMILAR TO RIKEN Q9BTI4 84% 323 1492 CDNA 8430408O15 GENE. 60% 353 421 HNTAF42 824082 2265 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 63% 380 673 CLONE KAT08285. HNTCG32 897776 2266 HMMER PFAM: Sodium/hydrogen exchanger PF00999 244.7 178 789 2.1.1 family WUblastx.64 (AAK54508) Nonselective sodium AAK54508 60% 13 837 potassium/proton exc 52% 772 1089 HNTNY89 886188 2267 WUblastx.64 (Q9H400) DJ583P15.4.1 (NOVEL Q9H400 90% 338 1006 PROTEIN (TRANSLATION OF 100% 158 187 CDNA FLJ20406 (E 68% 307 354 HNTRQ40 809100 2269 WUblastx.64 (Q9UBC2) EPIDERMAL GROWTH Q9UBC2 64% 1498 1857 FACTOR RECEPTOR SUBSTRATE 76% 11 451 EPS15R. 92% 492 1604 66% 1810 1845 57% 1567 1608 36% 32 178 40% 202 486 40% 1540 1623 48% 1567 1686 41% 1398 1544 33% 1395 1589 50% 1452 1544 34% 1377 1547 36% 495 695 35% 20 196 40% 519 623 23% 750 1610 HOAAJ09 654862 2273 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 82% 560 510 FOR MGC: 5149). 77% 769 572 HOAAL10 566867 2274 WUblastx.64 (Q9P1N7) PRO0974. Q9P1N7 42% 1113 1217 58% 929 1129 HOABC12 801922 2276 WUblastx.64 (Q13391) HYPOTHETICAL Q13391 100% 208 405 PROTEIN 384D8_6. 97% 5 208 HOABH36 740758 2277 WUblastx.64 (Q9BSY5) UNKNOWN (PROTEIN Q9BSY5 94% 147 1073 FOR IMAGE: 3831362) 86% 1078 1353 (FRAGMENT). 83% 10 177 HOBNA89 604999 2278 WUblastx.64 microtubule-associated pir|A54602|A54602 51% 369 7 serine/threonine protein kinase MAST205 - mouse HOBNF51 580861 2279 WUblastx.64 (O00410) IMPORTIN BETA-3 IMB3_HUMAN 95% 10 414 SUBUNIT (KARYOPHERIN BETA-3 20% 271 375 SUBUNI 99% 395 1051 HODAH24 656887 2280 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 68% 1326 1189 FOR MGC: 5149). HODAH46 862551 2281 WUblastx.64 (Q9H6Y7) CDNA: FLJ21676 FIS, Q9H6Y7 82% 264 608 CLONE COL09164. HODAV25 787471 2282 WUblastx.64 (Q9UHS9) PRO1914 PROTEIN. Q9UHS9 100% 72 1 HODAW64 840069 2283 WUblastx.64 (Q9H7T2) CDNA FLJ14295 FIS, Q9H7T2 93% 634 1263 CLONE PLACE1008426, WEAKLY SIMILAR TO RES HODAY17 806341 2284 WUblastx.64 (Q9H7L0) FLJ00062 PROTEIN Q9H7L0 90% 492 1478 (FRAGMENT). 100% 77 487 HODBF86 605001 2289 WUblastx.64 (Q9BRM8) UNKNOWN (PROTEIN Q9BRM8 43% 189 410 FOR MGC: 13219). HODDJ25 824592 2299 WUblastx.64 (Q9C0K7) AMYOTROPHIC Q9C0K7 100% 266 1519 LATERAL SCLEROSIS 2. HODDQ06 834824 2302 WUblastx.64 (Q9NZK6) PDZ-BINDING KINASE. Q9NZK6 94% 200 1165 HODEA20 840376 2303 WUblastx.64 (Q9BXY9) RALBP1. Q9BXY9 84% 141 1229 68% 6 179 HOEBI94 795312 2306 WUblastx.64 (Q9Y4J5) RIBONUCLEOPROTEIN Q9Y4J5 74% 823 999 (HNRNP 2H9). 100% 695 820 33% 710 820 100% 167 469 HOEBJ70 836143 2307 WUblastx.64 (AAH09229) Unknown (protein for AAH09229 100% 560 646 MGC: 16480). HOECB33 840378 2308 WUblastx.64 (Q9H6D8) CDNA: FLJ22362 FIS, Q9H6D8 100% 10 525 CLONE HRC06544. HOECX21 842866 2309 WUblastx.64 (Q9HC05) CD003 PROTEIN. Q9HC05 100% 368 225 HOEDE27 825262 2310 WUblastx.64 (Q9P195) PRO1722. Q9P195 55% 914 759 67% 780 649 HOEEK81 844390 2311 WUblastx.64 (BAB22091) Adult male kidney BAB22091 60% 51 596 cDNA, RIKEN full-lengt HOEEZ62 638833 2312 WUblastx.64 (Q9T9V8) NADH Q9T9V8 88% 66 173 DEHYDROGENASE SUBUNIT 3. 83% 303 338 HOEFJ26 847422 2313 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 77% 1198 995 CLONE LNG09295. HOFMF63 847423 2315 WUblastx.64 L6 surface protein - human pir|A42926|A42926 82% 409 690 77% 86 409 HOFMJ65 664498 2316 WUblastx.64 (P70560) COLLAGEN ALPHA 1(XII) CA1C_RAT 74% 987 1160 CHAIN (FRAGMENT). HOFMO16 596835 2318 HMMER PFAM: 3-beta hydroxysteroid PF01073 68.3 149 346 2.1.1 dehydrogenase/isomerase family WUblastx.64 (Q9BSN9) 3 BETA-HYDROXY- Q9BSN9 75% 11 184 DELTA 5-C27-STEROID 83% 349 438 OXIDOREDUCTASE. 94% 290 346 HOFMV22 812864 2321 WUblastx.64 (Q9P039) HSPC113. Q9P039 60% 180 293 50% 4 198 HOFNY15 668259 2323 WUblastx.64 (Q9H9J2) CDNA FLJ12701 FIS, Q9H9J2 73% 69 326 CLONE NT2RP1000730. 83% 331 999 HOFNY28 603911 2324 WUblastx.64 interferon gamma receptor accessory pir|I38500|I38500 59% 13 654 factor-1 precursor - human HOGAA41 843499 2326 WUblastx.64 (Q9D3B1) 6330408J20RIK Q9D3B1 96% 580 1257 PROTEIN. HOGAB51 825278 2327 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 74% 919 1119 CLONE LNG09295. HOGAH40 790978 2328 WUblastx.64 (Q9NX63) CDNA FLJ20420 FIS, Q9NX63 93% 185 844 CLONE KAT02462. HOGAP06 823364 2329 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 70% 1445 1669 PROTEIN. HOGAR71 722242 2331 HMMER PFAM: PMP-22/EMP/MP20/Claudin PF00822 336 96 632 2.1.1 family WUblastx.64 (O95471) CLAUDIN-7. CLD7_HUMAN 100% 87 719 HOGCC26 834792 2332 WUblastx.64 (Q9HCN4) XPA BINDING PROTEIN Q9HCN4 87% 19 1140 1. HOGCD78 610217 2333 WUblastx.64 (Q9H6Q7) CDNA: FLJ21979 FIS, Q9H6Q7 95% 428 487 CLONE HEP06065 (FRAGMENT). 100% 217 321 HOGCK03 847427 2334 WUblastx.64 (Q99LE1) UNKNOWN (PROTEIN Q99LE1 53% 310 405 FOR MGC: 7036). 78% 369 860 HOGCL01 846353 2335 HMMER PFAM: KOW motif PF00467 40.2 332 442 2.1.1 WUblastx.64 (Q96A35) Similar to mitochondrial Q96A35 100% 167 814 ribosomal protein L24. HOHBB36 847428 2336 WUblastx.64 myosin I alpha chain - mouse pir|A45438|A45438 90% 2093 3 93% 2237 2103 HOHBC57 813392 2337 WUblastx.64 (Q9VEL9) CG4090 PROTEIN. Q9VEL9 21% 2162 1767 22% 4081 3632 31% 1961 1836 18% 2582 1737 17% 4772 4509 20% 1793 1374 20% 2582 2376 26% 1897 1652 21% 3583 2564 22% 3550 2534 21% 2087 1809 24% 1970 1704 22% 4084 2534 38% 575 498 20% 2021 1773 21% 3556 2534 HOHBO66 853375 2338 WUblastx.64 (Q9NY61) DED PROTEIN Q9NY61 94% 1322 828 (APOPTOSIS ANTAGONIZING 88% 844 212 TRANSCRIPTION FACTOR). HONAH67 821419 2342 WUblastx.64 (Q9H7T6) CDNA FLJ14272 FIS, Q9H7T6 97% 1192 1338 CLONE PLACE1004793, WEAKLY 96% 513 1172 SIMILAR TO RET HOOAC84 604043 2343 WUblastx.64 (Q9UPN3) ACTIN CROSS-LINKING ACF7_HUMAN 84% 1324 1482 FAMILY PROTEIN 7 100% 325 444 (MACROPHIN) ( 100% 592 690 29% 1381 1461 36% 1387 1461 HOPBP13 825243 2344 WUblastx.64 (Q9P195) PRO1722. Q9P195 63% 2105 1851 HORBI80 877660 2346 WUblastx.64 (Q9NY33) DIPEPTIDYL- DPP3_HUMAN 100% 30 92 PEPTIDASE III (EC 3.4.14.4) (DPP 99% 92 2239 III) ( HORBL77 852099 2347 WUblastx.64 (Q9CWX4) 2410001E19RIK Q9CWX4 27% 852 1034 PROTEIN. 31% 212 784 HOSEM81 846354 2350 WUblastx.64 (O75872) RAB3-GAP O75872 100% 1012 803 REGULATORY DOMAIN. HOSEO83 847083 2351 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 68% 356 412 CLONE KAIA0536. 73% 310 354 53% 551 769 HOSFR35 845994 2352 WUblastx.64 (Q9NXV7) CDNA FLJ20035 FIS, Q9NXV7 63% 9 320 CLONE COL00213. 67% 316 780 HOUBC29 872565 2354 WUblastx.64 GTP-binding protein rab2 - rat pir|B39963|B39963 94% 763 876 HOUBG39 601696 2355 WUblastx.64 (Q9H6G8) CDNA: FLJ22294 FIS, Q9H6G8 79% 1603 1403 CLONE HRC04426. HOUCD12 605006 2356 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 69% 250 555 CLONE COL04765. HOUDB17 603417 2357 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 66% 672 367 PRODUCT. HOUIG68 793749 2363 WUblastx.64 (Q9NVC9) CDNA FLJ10808 FIS, Q9NVC9 100% 2 355 CLONE NT2RP4000879, WEAKLY SIMILAR TO UBI HOVAJ68 741098 2369 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 55% 1588 1421 CLONE MAMMA1000472. HOVAW46 892162 2370 WUblastx.64 (O00363) PUTATIVE P150. O00363 41% 395 345 39% 1030 773 37% 1359 1009 71% 759 697 45% 697 359 HOVBB19 843768 2371 WUblastx.64 catalase (EC 1.11.1.6) - pir|I40767|I40767 97% 307 197 Campylobacter jejuni HOVBI16 581097 2374 WUblastx.64 (AAG50170) Tripartite motif protein AAG50170 63% 83 115 TRIM28 alpha. 83% 202 702 HOVCO53 564632 2380 WUblastx.64 (Q9H8N2) CDNA FLJ13381 FIS, Q9H8N2 52% 1321 1127 CLONE PLACE1001010. HPBDE33 853660 2384 WUblastx.64 hypothetical protein pir|T12458|T12458 64% 257 985 DKFZp564O0823.1 - human HPBDE33 897372 2385 WUblastx.64 hypothetical protein pir|T12458|T12458 65% 257 985 DKFZp564O0823.1 - human HPCAG17 757836 2387 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 64% 1306 1013 CLONE COL03536. HPCAG17 863943 2388 WUblastx.64 (Q9H387) PRO2550. Q9H387 71% 1141 1007 71% 1008 850 HPDDQ28 566793 2391 WUblastx.64 (O95357) PUTATIVE G PROTEIN- O95357 98% 231 440 COUPLED RECEPTOR (CDNA 100% 10 231 FLJ10899 FIS, CLON HPDDT14 580867 2392 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 71% 193 5 PROTEIN. HPFCP75 581083 2399 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 53% 656 949 CLONE KAT08285. HPFDB66 829318 2400 WUblastx.64 (Q9NTM9) BA483F11.3 (CGI-32 Q9NTM9 100% 102 920 PROTEIN). HPFDD28 824856 2401 WUblastx.64 (Q9P195) PRO1722. Q9P195 66% 672 628 70% 622 551 69% 815 660 HPIAK27 852658 2404 WUblastx.64 (AAH06621) RIKEN cDNA AAH06621 83% 16 1050 2810403A07 gene. 70% 1286 1864 95% 1162 1293 40% 1162 1290 34% 1201 1269 44% 1174 1245 HPIAL55 847429 2405 WUblastx.64 (Q9D0W4) 1110060O18RIK Q9D0W4 96% 506 592 PROTEIN. 90% 1088 1150 72% 694 1002 100% 82 231 90% 1778 1807 HPIAT18 877663 2406 WUblastx.64 (Q9H397) PRO2852. Q9H397 84% 3823 3728 74% 3735 3571 HPIAZ52 801924 2407 WUblastx.64 (Q9H387) PRO2550. Q9H387 68% 2082 1987 67% 1988 1833 HPIBA07 886191 2408 WUblastx.64 (Q99ML9) ARKADIA. Q99ML9 73% 61 1482 HPIBA24 840379 2409 WUblastx.64 (Q9ULZ5) GONADOTROPIN Q9ULZ5 50% 27 1865 INDUCIBLE TRANSCRIPTION 56% 378 1793 REPRESSOR-4. 57% 420 1805 53% 549 1868 53% 330 1544 HPIBI40 588944 2410 WUblastx.64 (O43264) ZW10_HUMAN 99% 1 339 CENTROMERE/KINETOCHORE 98% 336 860 PROTEIN ZW10 HOMOLOG. HPJAB75 841653 2413 WUblastx.64 (Q9BZ63) FKSG60. Q9BZ63 80% 1581 1273 HPJAN76 826185 2414 WUblastx.64 (O43194) PUTATIVE G PROTEIN- GP39_HUMAN 89% 328 828 COUPLED RECEPTOR GPR39. HPJAN76 854893 2415 WUblastx.64 (O43194) PUTATIVE G PROTEIN- GP39_HUMAN 89% 328 828 COUPLED RECEPTOR GPR39. HPJAU94 826719 2416 WUblastx.64 (O00371) L1 ELEMENT L1.21 P40 O00371 83% 3944 4159 PROTEIN. 56% 3529 3921 HPJAW78 812766 2417 WUblastx.64 (Q9D1W2) C030013D06RIK Q9D1W2 44% 87 254 PROTEIN. 38% 96 266 40% 77 262 45% 87 260 47% 86 256 43% 243 85 40% 259 86 39% 249 67 41% 261 88 HPJBS16 608307 2418 WUblastx.64 (CAC39435) Epigen protein precursor. CAC39435 64% 31 357 HPJBU04 798101 2419 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 72% 830 567 CLONE COL03536. 47% 2315 2265 HPJCP75 886192 2421 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 61% 1459 1704 PROTEIN. HPJCV35 827317 2422 WUblastx.64 (Q9Y5S2) CDC42-BINDING Q9Y5S2 92% 2685 2951 PROTEIN KINASE BETA. 100% 533 610 HPJCX13 852869 2423 HMMER PFAM: Reverse transcriptase (RNA- PF00078 307.8 3321 4148 2.1.1 dependent DNA polymerase) WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 96% 4105 266 human HPMBT05 658715 2427 WUblastx.64 (Q14287) HYPOTHETICAL Q14287 73% 491 592 PROTEIN (FRAGMENT). 85% 403 483 HPMDD27 830748 2432 WUblastx.64 (Q9NZX0) HSPC068. Q9NZX0 96% 2 1417 HPMDF45 638148 2433 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 67% 1678 1857 CLONE KAIA0536. 72% 1856 1987 HPMDP57 567303 2434 WUblastx.64 (Q9H387) PRO2550. Q9H387 64% 1286 1074 77% 1447 1211 HPMEG72 795709 2435 WUblastx.64 (O00549) ORF2-LIKE PROTEIN O00549 55% 1844 2056 (FRAGMENT). HPMFM70 756931 2436 WUblastx.64 (Q9H387) PRO2550. Q9H387 81% 2108 1998 61% 2286 2248 68% 2248 2105 HPMFP48 597457 2437 WUblastx.64 (Q9P195) PRO1722. Q9P195 66% 987 853 69% 1130 975 HPMFW01 844865 2438 WUblastx.64 (Q9NP48) PUTATIVE LIPID Q9NP48 97% 973 1113 KINASE (CDNA FLJ10842 FIS, CLONE NT2RP4001343 HPMGW43 830452 2440 WUblastx.64 vesicle-associated membrane protein- pir|JG0186|JG0186 100% 31 72 associated protein B - human 95% 59 427 HPMKB09 900362 2442 WUblastx.64 (Q9UJU6) SRC HOMOLOGY 3 Q9UJU6 92% 798 1406 DOMAIN-CONTAINING PROTEIN 92% 117 656 HIP-55 (DREBRIN F). HPMSH26 780109 2443 WUblastx.64 (Q9NQC3) NOGO-A PROTEIN. Q9NQC3 69% 1592 1014 70% 1748 1578 HPMSH96 610023 2444 WUblastx.64 (AAH00127) Glutathione-S- AAH00127 88% 1203 1505 transferase like, glutathi HPQAJ27 782957 2446 WUblastx.64 peptidyl-prolyl cis-trans isomerase pir|B81216|B81216 54% 1370 987 NMB0281 [imported] - Neisseria 77% 973 707 meningitidis (strain MC58 serogroup B) HPQAN50 788813 2447 WUblastx.64 (O00549) ORF2-LIKE PROTEIN O00549 35% 381 10 (FRAGMENT). 32% 1151 222 38% 1285 590 HPRAD30 805969 2458 WUblastx.64 (Q9P1C6) PRO2738. Q9P1C6 52% 2465 2265 HPRCC91 638151 2459 WUblastx.64 (Q9H1R7) BA534G20.4 Q9H1R7 100% 2 97 (SUPERVILLIN) (FRAGMENT). HPRCF40 834808 2460 WUblastx.64 (AAH07482) Similar to conserved AAH07482 97% 3 221 membrane protein at HPRCF50 638153 2461 WUblastx.64 (Q9H387) PRO2550. Q9H387 66% 1787 1912 58% 1614 1790 HPRCM72 813512 2463 WUblastx.64 (Q9D3K9) 2810468K05RIK Q9D3K9 45% 296 526 PROTEIN. HPRCS59 601523 2464 WUblastx.64 (Q61787) ORF 2. Q61787 30% 746 189 HPRCT73 610024 2465 WUblastx.64 (Q9NPZ5) B3G2_HUMAN 96% 1231 1145 GALACTOSYLGALACTOSYLXYL 89% 1476 1300 OSYLPROTEIN 3-BETA- GLUCURON HPTRE80 884167 2467 WUblastx.64 (O43819) SCO2 PROTEIN SCO2_HUMAN 85% 779 39 HOMOLOG PRECURSOR. HPTRI42 655362 2468 WUblastx.64 (Q9BVA7) UNKNOWN (PROTEIN Q9BVA7 85% 3 611 FOR MGC: 5621). HPTTT62 561954 2473 WUblastx.64 (Q9H3X5) HYPOTHETICAL 85.5 KDA Q9H3X5 100% 8 169 PROTEIN (FRAGMENT). HPTVH24 831983 2474 WUblastx.64 (Q9H7Z7) CDNA FLJ14038 FIS, Q9H7Z7 90% 3 974 CLONE HEMBA1005206. HPTVI96 636064 2477 WUblastx.64 (Q9H5X3) HYPOTHETICAL 13.8 KDA Q9H5X3 77% 42 416 PROTEIN. HPVAA15 783074 2478 WUblastx.64 (AAK54355) ATP-binding cassette AAK54355 40% 4 483 transporter family 33% 7 477 31% 487 870 30% 487 828 HPWAS27 536008 2484 WUblastx.64 (Q9P0E3) HSPC093 (FRAGMENT). Q9P0E3 40% 1042 1146 65% 512 625 HPWAV82 830084 2486 WUblastx.64 (Q9H387) PRO2550. Q9H387 79% 3 89 69% 104 229 HPWBA36 840380 2487 WUblastx.64 (Q9H387) PRO2550. Q9H387 54% 1183 1073 51% 1238 1152 62% 1380 1219 HPWTF23 843700 2488 HMMER PFAM: TSC-22/dip/bun family PF01166 146.4 442 621 2.1.1 WUblastx.64 (Q99576) GLUCOCORTICOID- GILZ_HUMAN 94% 271 672 INDUCED LEUCINE ZIPPER PROTEIN (DEL HPWTF23 844775 2489 HMMER PFAM: TSC-22/dip/bun family PF01166 146.4 442 621 2.1.1 WUblastx.64 (Q99576) GLUCOCORTICOID- GILZ_HUMAN 94% 271 672 INDUCED LEUCINE ZIPPER PROTEIN (DEL HPWTF53 844737 2490 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 48% 1855 2082 PROTEIN. HRAAC36 798104 2494 WUblastx.64 (Q26195) PVA1 GENE. Q26195 65% 835 758 62% 750 565 41% 2229 2119 HRAAF59 847086 2495 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 56% 1519 1247 CLONE COL04765. HRAAG89 621271 2496 WUblastx.64 (Q9GMW5) HYPOTHETICAL 45.1 KDA Q9GMW5 93% 2 187 PROTEIN. HRAAZ12 834637 2498 WUblastx.64 (AAG41897) Neuropilin-2a(17). AAG41897 100% 1361 1435 92% 863 901 56% 385 579 96% 579 878 79% 901 1356 HRABP28 823344 2500 WUblastx.64 (AAK55521) PRO0764. AAK55521 44% 1136 975 44% 1245 1111 HRABU56 621381 2501 WUblastx.64 (O75876) RNA-BINDING PROTEIN. O75876 90% 105 320 HRABZ80 562230 2502 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 64% 1166 1116 CLONE COL03536. 54% 1104 1000 HRACB01 637647 2503 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 52% 1423 1349 PROTEIN. 52% 1344 1135 HRACI39 840461 2504 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 59% 1346 1146 PRODUCT. HRADU15 801926 2505 WUblastx.64 (Q9H7S6) CDNA FLJ14310 FIS, Q9H7S6 75% 645 586 CLONE PLACE3000271. 74% 787 638 HRDAH04 651356 2506 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 38% 1336 1229 CLONE COL04765. 60% 1478 1329 HRDBA20 637709 2507 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 80% 1113 1069 CLONE MAMMA1000472. 71% 1049 1008 79% 1267 1106 HRDBD32 637710 2508 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 80% 1113 1069 CLONE MAMMA1000472. 71% 1049 1008 79% 1267 1106 HRDBL01 631169 2509 WUblastx.64 (Q9P195) PRO1722. Q9P195 65% 934 755 HRDDM85 799542 2510 WUblastx.64 (O95662) POT. ORF VI O95662 60% 1034 1096 (FRAGMENT). 56% 538 1077 HRDEJ86 695755 2512 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 74% 827 627 CLONE KAIA0536. HRDFE30 750872 2514 WUblastx.64 (Q9NWU2) BA305P22.1. Q9NWU2 89% 533 823 95% 141 548 HRDFT83 606799 2515 WUblastx.64 (O60448) NEURONAL THREAD O60448 60% 442 377 PROTEIN AD7C-NTP. 84% 584 528 60% 439 380 53% 578 459 61% 932 768 30% 778 641 68% 779 714 70% 942 769 58% 387 337 47% 948 892 42% 584 294 100% 691 668 62% 948 664 HRGCA01 589970 2516 WUblastx.64 (Q9H387) PRO2550. Q9H387 57% 1318 1016 HRGCA06 866189 2517 HMMER PFAM: Ribosomal protein S16 PF00886 69.1 143 325 2.1.1 WUblastx.64 (Q9Y3D3) 28S RIBOSOMAL RT16_HUMAN 99% 74 445 PROTEIN S16, MITOCHONDRIAL PRECURSOR HRGSE38 898233 2518 WUblastx.64 ATPase inhibitor precursor, pir|JC7175|JC7175 83% 77 367 mitochondrial - human HRLME03 610614 2520 WUblastx.64 (Q9H3B9) PRO0956. Q9H3B9 39% 220 122 68% 111 46 HROAP64 835467 2522 WUblastx.64 (Q24333) ELASTIN LIKE PROTEIN Q24333 100% 46 117 (FRAGMENT). HROAS35 827304 2523 WUblastx.64 cytochrome-c oxidase (EC 1.9.3.1) pir|A00482|OTHU3 77% 483 881 chain III - human 1 HROBJ10 836368 2525 WUblastx.64 (Q9Y5P3) RETINOIC ACID- RAI2_HUMAN 36% 1719 1441 INDUCED PROTEIN 2. 96% 1817 1719 96% 1054 671 100% 1445 1044 HRTAE88 822964 2529 WUblastx.64 (Q13579) MARINER Q13579 76% 2579 2845 TRANSPOSASE. HRTAP63 780698 2530 WUblastx.64 (Q9Y3C9) CGI-127 PROTEIN. Q9Y3C9 100% 498 860 HSAAN03 599334 2532 WUblastx.64 (Q9H387) PRO2550. Q9H387 70% 924 832 72% 676 602 77% 1094 921 HSAAS05 703244 2533 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 55% 1475 1416 PRODUCT. 55% 1419 1216 HSAAW13 821334 2534 WUblastx.64 (AAK57641) CAMP-specific cyclic AAK57641 99% 1 1101 nucleotide phosphod HSATA61 801927 2538 WUblastx.64 (Q9CR30) 1110007C05RIK Q9CR30 73% 6 224 PROTEIN. HSATG66 824903 2539 WUblastx.64 (Q9JKP5) MUSCLEBLIND. Q9JKP5 100% 561 590 52% 13 213 83% 1 465 HSATI91 838829 2540 WUblastx.64 hypothetical protein (L1H 3′ region) - pir|B34087|B34087 43% 345 127 human 35% 808 515 30% 828 580 50% 527 375 32% 128 45 HSATR50 826398 2541 WUblastx.64 (AAH02742) U6 snRNA-associated AAH02742 100% 397 483 Sm-like protein LSm8 HSATT82 566784 2542 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 57% 1004 891 CLONE HEMBA1006036. 68% 915 766 HSATW19 637658 2543 WUblastx.64 (Q9UI50) PRO0657 (FRAGMENT). Q9UI50 75% 448 669 HSATW67 604938 2544 WUblastx.64 pro-pol-dUTPase polyprotein - murine pir|T29097|T29097 63% 653 441 endogenous retrovirus ERV-L 82% 436 119 (fragment) HSATZ02 490895 2545 WUblastx.64 (Q9H387) PRO2550. Q9H387 75% 964 857 76% 858 742 HSAUB89 600369 2547 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 62% 359 631 PROTEIN. HSAUI53 866191 2548 WUblastx.64 retrovirus-related reverse transcriptase pir|B25313|GNLRL1 42% 362 9 pseudogene - slow loris 32% 562 365 44% 942 868 44% 1037 957 38% 570 436 37% 827 723 HSAUV74 866193 2549 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 62% 827 693 HSAUX39 823161 2550 WUblastx.64 (Q62510) ZINC FINGER PROTEIN Q62510 33% 960 1040 62 (FRAGMENT). 41% 1319 1645 43% 1244 1975 50% 1343 1975 52% 1349 1894 59% 1412 1975 45% 1235 1975 63% 1385 1972 52% 1349 1966 63% 1388 1966 48% 1130 1810 49% 1244 1975 64% 1367 1972 56% 1223 1975 HSAVE52 600370 2552 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 55% 1425 1366 CLONE COL04765. 56% 1384 1127 HSAVH32 603367 2553 WUblastx.64 (AAK55521) PRO0764. AAK55521 73% 811 767 56% 789 547 HSAVO11 566467 2555 WUblastx.64 (O60448) NEURONAL THREAD O60448 41% 1599 1207 PROTEIN AD7C-NTP. 66% 1597 1439 54% 1257 1192 64% 1582 1301 HSAVO17 738007 2556 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 72% 1407 1129 CLONE COL03536. HSAVQ13 606818 2557 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 66% 494 538 PROTEIN. 72% 406 471 HSAVR85 786185 2558 WUblastx.64 (Q9H396) PRO2870. Q9H396 100% 3791 3546 HSAVY92 606808 2559 WUblastx.64 (Q9H5T7) CDNA: FLJ23054 FIS, Q9H5T7 100% 19 123 CLONE LNG03193. HSAVZ05 690151 2560 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 65% 57 257 CLONE COL03536. 56% 256 303 67% 1529 1212 HSAWB58 738018 2561 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 75% 1386 1607 PROTEIN. HSAWH36 581087 2562 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 71% 1109 867 CLONE KAT08285. HSAWM20 606840 2563 WUblastx.64 (Q9P147) PRO2822. Q9P147 67% 488 387 57% 363 322 73% 591 490 HSAWM74 866195 2564 WUblastx.64 (O95166) MM46 (HT004 PROTEIN) O95166 100% 8 85 (MAP1 LIGHT CHAIN 3 RELATED PROTEIN). HSAWX70 872498 2565 WUblastx.64 (O00369) L1 ELEMENT L1.20 P40 O00369 43% 1309 1088 PROTEIN. 73% 1028 300 HSAXI10 598726 2567 WUblastx.64 (Q9H387) PRO2550. Q9H387 79% 643 512 75% 811 776 71% 776 621 HSAXL49 606826 2568 WUblastx.64 (Q9H387) PRO2550. Q9H387 63% 742 572 83% 863 717 HSAXL82 566801 2569 WUblastx.64 (Q9CXK1) 5730406I15RIK Q9CXK1 96% 533 613 PROTEIN. HSAXS06 668260 2572 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 50% 955 824 CLONE COL03536. 62% 1121 954 HSAYL24 608642 2574 WUblastx.64 (Q9UI59) PRO0478 PROTEIN. Q9UI59 73% 132 7 HSBAJ47 842694 2578 WUblastx.64 (Q9H189) SPHINGOSINE-1- Q9H189 100% 1 237 PHOSPHATASE. HSDDC55 663277 2580 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 82% 743 793 CLONE KAIA0536. 57% 490 741 HSDEA26 822812 2581 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 76% 1325 1387 75% 1140 1322 HSDGH56 853379 2587 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 94% 865 815 CLONE COL04765. 61% 1096 1058 70% 1058 879 HSDGM01 608310 2588 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 85% 722 781 PROTEIN. 53% 529 726 HSDGM42 870143 2589 WUblastx.64 (Q9BT21) SIMILAR TO RIKEN Q9BT21 92% 360 2639 CDNA 2610005L19 GENE (FRAGMENT). HSDGM42 824916 2590 WUblastx.64 (Q9BT21) SIMILAR TO RIKEN Q9BT21 91% 359 2638 CDNA 2610005L19 GENE (FRAGMENT). HSDGM42 852473 2591 WUblastx.64 (Q9BT21) SIMILAR TO RIKEN Q9BT21 95% 1544 606 CDNA 2610005L19 GENE (FRAGMENT). HSDGM42 861925 2592 WUblastx.64 (Q9BT21) SIMILAR TO RIKEN Q9BT21 92% 420 2699 CDNA 2610005L19 GENE (FRAGMENT). HSDGM42 865828 2593 WUblastx.64 (Q9BT21) SIMILAR TO RIKEN Q9BT21 92% 420 2699 CDNA 2610005L19 GENE (FRAGMENT). HSDGM42 886748 2594 WUblastx.64 (Q9BT21) SIMILAR TO RIKEN Q9BT21 92% 420 2699 CDNA 2610005L19 GENE (FRAGMENT). HSDIK31 847087 2597 WUblastx.64 (O15121) PUTATIVE FATTY ACID O15121 100% 19 369 DESATURASE MLD 97% 354 881 (DEGENERATIVE SPERMATOCYT HSDJC96 753284 2599 WUblastx.64 (Q9Y6A3) HYPOTHETICAL 5.6 KDA Q9Y6A3 100% 1946 1800 PROTEIN (FRAGMENT). HSDJF04 695756 2601 WUblastx.64 (O95890) UNKNOWN. O95890 94% 2 382 HSDJG47 847325 2602 WUblastx.64 (Q9H6U6) CDNA: FLJ21857 FIS, Q9H6U6 84% 59 1180 CLONE HEP02294. HSDJH72 805971 2603 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 66% 1500 1447 FOR MGC: 5149). 71% 1709 1485 HSDJL07 895387 2604 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 64% 764 898 CLONE LNG09295. 82% 1481 1290 HSDJR49 741079 2605 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 63% 1000 1122 CLONE COL04765. 70% 893 1015 68% 786 872 62% 1124 1210 73% 1000 1044 67% 569 733 HSDJV24 806246 2606 WUblastx.64 (Q9CWT1) 2410004N11RIK Q9CWT1 72% 539 1429 PROTEIN. HSDJV40 623716 2607 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 69% 1105 977 CLONE KAIA0536. 59% 1360 1295 38% 1583 1401 60% 1277 1104 HSDKA64 600372 2608 WUblastx.64 (Q9H387) PRO2550. Q9H387 80% 1340 1278 72% 1548 1342 HSDKF96 839730 2610 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 63% 1672 1412 PRODUCT. HSDZO08 827460 2611 WUblastx.64 (Q9UES6) I-1 RECEPTOR Q9UES6 74% 1 393 CANDIDATE PROTEIN. 30% 1 156 92% 449 1648 HSEBB18 877813 2613 WUblastx.64 (Q9NXX4) CDNA FLJ20005 FIS, Q9NXX4 89% 476 898 CLONE ADKA02526. HSFAM19 691371 2614 WUblastx.64 (O18966) EAG CHANNEL. O18966 83% 76 510 HSHAG54 834781 2615 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 51% 1028 738 CLONE LNG09295. HSHAS72 835991 2616 HMMER PFAM: G-protein gamma subunit. PF00631 139.1 225 422 2.1.1 WUblastx.64 (AAK53385) G protein gamma 12 AAK53385 100% 207 422 subunit. HSHAX04 812178 2617 WUblastx.64 peptidylprolyl isomerase (EC 5.2.1.8) pir|S66681|S66681 96% 14 916 A - human HSHBT15 581088 2618 WUblastx.64 (O95863) ZINC FINGER PROTEIN SNAI_HUMAN 100% 570 680 SNAI1 (SNAIL PROTEIN 79% 10 570 HOMOLOG) HSHCE85 855969 2619 WUblastx.64 (Q9Y546) DJ167A19.4 (NOVEL Q9Y546 100% 202 1224 PROTEIN). HSIAC81 783076 2620 WUblastx.64 (Q9H3T4) KLOTHO-RELATED Q9H3T4 99% 8 1261 PROTEIN 1. HSIAP01 897538 2622 WUblastx.64 (Q9NXP8) CDNA FLJ20124 FIS, Q9NXP8 92% 590 673 CLONE COL06056. 28% 218 535 100% 685 732 89% 71 613 HSIDZ25 658721 2625 WUblastx.64 (Q9HBN2) HYPOTHETICAL 15.8 KDA Q9HBN2 37% 1366 1280 PROTEIN. 80% 1689 1597 HSIEB64 651359 2626 WUblastx.64 (Q9P1J1) PRO1546. Q9P1J1 70% 1718 1747 60% 1860 1949 53% 1741 1875 HSIFO61 845026 2628 WUblastx.64 (O95831) PROGRAMED CELL PCD8_HUMAN 96% 95 1933 DEATH PROTEIN 8, MITOCHONDRIAL PREC HSIFO61 852715 2629 WUblastx.64 (O95831) PROGRAMED CELL PCD8_HUMAN 96% 183 2021 DEATH PROTEIN 8, MITOCHONDRIAL PREC HSIGC63 877486 2630 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 58% 1734 1354 CLONE KAIA0536. HSIGM95 840382 2631 WUblastx.64 (Q9H397) PRO2852. Q9H397 80% 818 759 75% 754 707 77% 942 823 HSJAN83 825132 2633 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 86% 1063 977 CLONE HEMBA1006036. 63% 963 898 57% 827 771 87% 839 768 77% 960 841 HSJAQ10 853383 2634 WUblastx.64 (AAH05957) Solute carrier family 25 AAH05957 100% 11 187 (mitochondrial HSJAR59 846364 2635 WUblastx.64 (Q9NZS6) GLUCOCORTICOID Q9NZS6 100% 16 138 RECEPTOR AF-1 SPECIFIC ELONGATION FACTOR (FRA HSJAU93 702019 2636 WUblastx.64 (Q9BUK4) SIMILAR TO Q9BUK4 74% 123 728 HYPOTHETICAL PROTEIN FLJ10709. HSKHV81 841590 2646 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 60% 935 801 CLONE MAMMA1000472. 26% 308 219 HSKYR49 806548 2648 WUblastx.64 (Q9BTV7) UNKNOWN (PROTEIN Q9BTV7 100% 10 456 FOR IMAGE: 3357127) (FRAGMENT). HSKYU81 899335 2649 WUblastx.64 (Q9Y2W2) SH3 DOMAIN-BINDING Q9Y2W2 84% 950 1276 PROTEIN SNP70 (NPW38-BINDING 100% 536 565 PROTEIN NPWB 31% 240 365 72% 6 533 HSKYY92 853384 2650 WUblastx.64 (Q9P0E3) HSPC093 (FRAGMENT). Q9P0E3 57% 1455 1351 50% 1348 1193 HSLAB11 823825 2651 WUblastx.64 (Q9CVT0) 1700040C17RIK Q9CVT0 99% 915 1427 PROTEIN (FRAGMENT). 34% 313 453 89% 187 915 HSLAS96 740764 2652 WUblastx.64 (Q9H387) PRO2550. Q9H387 62% 1173 1075 73% 1378 1175 HSLAW59 637669 2653 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 52% 578 423 CLONE HEMBA1006036. 70% 427 287 HSLCH54 562018 2654 WUblastx.64 (P51805) PLEXIN A3 PRECURSOR PLX4_HUMAN 97% 445 570 (PLEXIN 4) (TRANSMEMBRANE 81% 9 446 PROT HSLCH57 899415 2655 WUblastx.64 SREBP cleavage activating protein - pir|T18526|T18526 92% 1408 2205 Chinese hamster 39% 658 774 47% 352 459 HSLCI86 737626 2656 WUblastx.64 estrogen sulfotransferase (EC 2.8.2.—) - pir|JC2229|JC2229 33% 493 555 human 100% 704 1090 HSLCS31 604046 2657 HMMER PFAM: PPR repeat PF01535 22.6 493 597 2.1.1 WUblastx.64 (Q9H9R0) CDNA FLJ12598 FIS, Q9H9R0 98% 454 828 CLONE NT2RM4001384. 100% 826 1047 HSLCS34 751324 2658 WUblastx.64 (Q9NVE5) CDNA FLJ10785 FIS, Q9NVE5 70% 40 576 CLONE NT2RP4000457, WEAKLY 86% 375 1007 SIMILAR TO UBI HSLCV16 772948 2659 WUblastx.64 (Q9H0J7) HYPOTHETICAL 53.4 KDA Q9H0J7 99% 1 552 PROTEIN. 87% 1537 1698 HSLDW54 853386 2660 WUblastx.64 probable pol polyprotein-related pir|S21348|S21348 52% 732 631 protein 4 - rat 44% 625 452 34% 1115 738 HSLEC18 722249 2661 WUblastx.64 (Q96RP7) Galbetal-3GalNAc 3′- Q96RP7 97% 60 1319 sulfotransferase. HSLEG59 637671 2662 WUblastx.64 (Q14287) HYPOTHETICAL Q14287 44% 968 807 PROTEIN (FRAGMENT). 20% 745 614 50% 1150 941 HSLFR59 853388 2663 WUblastx.64 hypothetical protein pir|T46471|T46471 86% 877 987 DKFZp434L0130.1 - human 93% 987 1481 HSLGD91 883491 2664 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 70% 764 817 CLONE KAIA0536. 72% 816 926 HSNAQ52 600402 2672 WUblastx.64 (Q9BGZ4) HYPOTHETICAL 11.6 KDA Q9BGZ4 85% 197 156 PROTEIN. 81% 142 95 HSNAW06 580248 2674 WUblastx.64 (Q9GMI2) HYPOTHETICAL 9.4 KDA Q9GMI2 61% 219 413 PROTEIN. HSNBQ36 784994 2678 WUblastx.64 (Q9NUU5) CDNA FLJ11128 FIS, Q9NUU5 98% 358 732 CLONE PLACE1006236. HSNBS39 617125 2679 WUblastx.64 retrovirus-related hypothetical protein pir|S23650|S23650 76% 313 263 II - human 1 47% 82 26 38% 275 93 HSOAT44 847357 2681 WUblastx.64 (Q9GZY9) CDNA: FLJ20877 FIS, Q9GZY9 100% 10 195 CLONE ADKA02965 (CDNA: FLJ20871 FIS, CLO HSOBH11 794000 2683 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 58% 657 743 PROTEIN. 59% 508 603 HSOBW65 778388 2685 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 64% 160 110 PROTEIN (FRAGMENT). 67% 636 142 HSPAA89 825099 2686 WUblastx.64 (P05142) PROLINE-RICH PROTEIN PRP2_MOUSE 42% 210 148 MP-2 PRECURSOR. 37% 1144 641 HSQBL20 780518 2696 WUblastx.64 (Q9UBQ5) MRNA OF MUSCLE Q9UBQ5 100% 183 836 SPECIFIC GENE M9, COMPLETE CDS (ARG134 PROTEI HSQCY74 886110 2698 WUblastx.64 (Q9H455) DJ383J4.4 (A NOVEL Q9H455 99% 10 1455 PROTEIN SIMILAR TO ASPARTYL-TRNA SYNTHETA HSRAA81 695759 2705 WUblastx.64 (CAC38441) DJ1033B10.5.1 (SAC2 CAC38441 90% 9 749 (suppressor of actin 33% 596 694 100% 727 1086 HSRAO56 719816 2706 WUblastx.64 (Q9HBS7) HYPOTHETICAL 14.2 KDA Q9HBS7 68% 1419 1324 PROTEIN. 72% 1592 1416 HSRAV28 581102 2707 WUblastx.64 (Q9H387) PRO2550. Q9H387 61% 655 524 61% 823 785 75% 788 654 HSRDW57 562019 2709 WUblastx.64 (BAB55068) CDNA FLJ14466 fis, BAB55068 100% 8 259 clone MAMMA1000416. 91% 262 495 83% 485 520 HSREC72 601368 2710 WUblastx.64 (Q9H387) PRO2550. Q9H387 88% 838 812 69% 783 625 HSREG42 839481 2711 WUblastx.64 (Q9VZZ4) PXN PROTEIN. Q9VZZ4 47% 6 1484 HSRFD18 840771 2712 WUblastx.64 (Q9H941) CDNA FLJ13033 FIS, Q9H941 100% 437 559 CLONE NT2RP3001126. HSRGZ11 801929 2713 WUblastx.64 (Q9BYN8) DJ534B8.3 (NOVEL Q9BYN8 100% 253 384 PROTEIN). HSRHB59 840384 2714 WUblastx.64 (Q9NWT0) HYPOTHETICAL 17.7 KDA Q9NWT0 100% 8 121 PROTEIN. 100% 123 308 HSSCC66 559402 2716 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 65% 548 667 PRODUCT. HSSEL28 838618 2721 WUblastx.64 (Q9P195) PRO1722. Q9P195 68% 755 1042 HSSFP88 658726 2722 HMMER PFAM: Zinc finger, C3HC4 type PF00097 36.1 828 953 2.1.1 (RING finger) WUblastx.64 (Q9H6Y7) CDNA: FLJ21676 FIS, Q9H6Y7 90% 204 1133 CLONE COL09164. HSSGS62 741162 2723 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 60% 899 762 PROTEIN. 60% 200 141 HSSJA23 847089 2724 WUblastx.64 (Q9BTH7) UNKNOWN (PROTEIN Q9BTH7 94% 272 685 FOR MGC: 5601). 96% 1379 1561 HSSJF26 567431 2725 WUblastx.64 (Q9HBS7) HYPOTHETICAL 14.2 KDA Q9HBS7 76% 600 689 PROTEIN. 46% 401 580 HSSJM47 796096 2727 WUblastx.64 (Q9UJ98) STROMAL ANTIGEN 3, Q9UJ98 77% 129 194 (STAG3). 75% 527 637 96% 899 994 HSSJW30 779822 2728 WUblastx.64 (BAB55147) CDNA FLJ14580 fis, BAB55147 86% 2 46 clone NT2RM4001204. 92% 45 488 HSSJW30 850566 2729 WUblastx.64 (BAB55147) CDNA FLJ14580 fis, BAB55147 67% 14 703 clone NT2RM4001204. HSSJW30 867721 2730 WUblastx.64 (BAB55147) CDNA FLJ14580 fis, BAB55147 82% 672 842 clone NT2RM4001204. 100% 625 675 89% 76 186 46% 131 220 60% 24 83 HSSMY35 740765 2731 WUblastx.64 (Q9H7J9) FLJ00075 PROTEIN Q9H7J9 96% 20 217 (FRAGMENT). HSTAL93 841863 2732 WUblastx.64 (AAK52433) Low density lipoprotein AAK52433 98% 287 799 receptor-related 35% 284 787 35% 299 787 35% 287 637 HSUAF06 863206 2734 WUblastx.64 pol polyprotein - Cas-Br-E murine pir|A26103|A26103 47% 630 944 leukemia virus (fragment) 50% 1194 1307 HSUBX67 751266 2735 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 61% 624 499 PRODUCT. 64% 766 608 HSUSB73 603912 2736 WUblastx.64 (Q9GMU5) HYPOTHETICAL 14.1 KDA Q9GMU5 57% 1158 1117 PROTEIN. 80% 130 56 HSVAC05 836051 2737 WUblastx.64 (Q9H6G8) CDNA: FLJ22294 FIS, Q9H6G8 64% 688 539 CLONE HRC04426. HSVBA83 812059 2742 WUblastx.64 (O60593) ARG/ABL-INTERACTING O60593 98% 313 516 PROTEIN ARGBP2B (FRAGMENT). HSVBY62 637113 2745 WUblastx.64 (Q9Y2Q7) HSPC005 PROTEIN Q9Y2Q7 100% 35 217 (C11ORF10) (CHROMOSOME 11 OPEN READING FRAME HSXAI44 590743 2750 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 90% 1024 992 CLONE COL04765. 73% 1007 828 HSXAS59 838072 2752 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 85% 1515 1474 CLONE COL04765. 81% 1303 1256 62% 1478 1305 HSXAY60 737753 2754 WUblastx.64 (Q9N032) UNNAMED PROTEIN Q9N032 59% 1742 1602 PRODUCT. HSXCA83 830046 2756 WUblastx.64 (Q9UHD2) TANK BINDING Q9UHD2 39% 1121 1219 KINASE TBK1 (NF-KB- 98% 1215 1382 ACTIVATING KINASE NAK). 93% 109 1197 HSXCX20 658728 2757 WUblastx.64 (Q9BXA5) G-PROTEIN COUPLED Q9BXA5 100% 891 1049 RECEPTOR 91. 86% 61 942 HSXFG21 805972 2758 WUblastx.64 (Q9CQQ2) 1700066F09RIK Q9CQQ2 47% 499 933 PROTEIN (FRAGMENT). HSXFH82 699863 2759 WUblastx.64 (Q9MZZ7) HYPOTHETICAL 16.3 KDA Q9MZZ7 69% 1447 1611 PROTEIN. 92% 1124 1162 65% 1146 1598 HSYBR79 873848 2761 WUblastx.64 vesicle-associated membrane protein- pir|JG0186|JG0186 100% 193 861 associated protein B - human HSYBV44 753253 2762 WUblastx.64 (Q9B2U5) ATP SYNTHASE 6. Q9B2U5 66% 84 761 HSYBZ94 799543 2763 WUblastx.64 (Q9BZ73) NIR2. Q9BZ73 98% 2168 2392 99% 2388 2924 33% 1313 1399 33% 1895 1999 79% 21 2174 HT3AB13 841680 2764 WUblastx.64 (Q9UP93) SHORT FORM Q9UP93 53% 663 980 TRANSCRIPTION FACTOR C-MAF. 74% 271 612 HT4SB02 837688 2765 HMMER PFAM: emp24/gp25L/p24 family PF01105 231.4 78 521 2.1.1 WUblastx.64 protein trafficking protein tmp21-I - pir|G01159|G01159 100% 84 524 human 100% 21 74 HT4SB81 756723 2767 WUblastx.64 (O95621) TIC. O95621 95% 3 194 77% 580 789 62% 185 631 41% 19 54 HT4SB81 844512 2768 WUblastx.64 (O95621) TIC. O95621 72% 185 790 95% 3 194 41% 19 54 57% 872 913 HT4SB81 858154 2769 WUblastx.64 (O95621) TIC. O95621 71% 185 790 41% 19 54 95% 3 194 57% 872 913 HTABF81 610040 2771 WUblastx.64 (Q9NRR3) NON-KINASE CDC42 Q9NRR3 100% 48 299 EFFECTOR PROTEIN SPEC2. HTACX63 602694 2772 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 65% 919 860 CLONE COL04765. 78% 825 784 53% 1067 921 HTADC63 842131 2773 WUblastx.64 (Q9D7U1) 2210407P13RIK Q9D7U1 40% 557 793 PROTEIN. 46% 162 557 HTADO61 695760 2774 WUblastx.64 hypothetical protein pir|T42648|T42648 100% 9 95 DKFZp434C1415.1 - human HTAEC59 846728 2776 WUblastx.64 ubiquitin-conjugating enzyme pir|S53358|S53358 100% 409 591 E2.17 kB - rat 100% 156 383 HTAED89 801931 2777 HMMER PFAM: 7 transmembrane receptor PF00003 78.6 1240 1578 2.1.1 (metabotropic glutamate family) WUblastx.64 (O35363) CALCIUM SENSING O35363 51% 1590 1718 RECEPTOR, RELATED SEQUENCE 38% 858 920 2 (CALCIUM-SENSIN 38% 908 1171 53% 1216 1611 HTAEO35 732379 2780 WUblastx.64 (Q9H387) PRO2550. Q9H387 73% 1476 1195 HTDAF68 637685 2781 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 72% 892 1041 CLONE COL04765. 50% 2 91 70% 711 890 HTDAI38 878930 2782 WUblastx.64 (Q9NX73) CDNA FLJ20400 FIS, Q9NX73 88% 954 1268 CLONE KAT00587 (FRAGMENT). 30% 361 669 34% 1146 1874 37% 1314 1898 100% 343 678 32% 930 1628 HTECE87 702025 2786 WUblastx.64 (Q9NVC3) CDNA FLJ10815 FIS, Q9NVC3 100% 53 133 CLONE NT2RP4000989, WEAKLY 100% 133 279 SIMILAR TO UNC HTEDF78 564215 2787 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 75% 1545 1423 PROTEIN. HTEDX05 862064 2789 WUblastx.64 (Q9D4H6) 4932415A06RIK Q9D4H6 79% 16 441 PROTEIN. 86% 429 1649 HTEEC19 862065 2790 WUblastx.64 translation initiation factor IF-2 pir|T43483|T43483 100% 11 1321 homolog [similarity] - 1 HTEGH03 815562 2791 WUblastx.64 (Q9UHB5) EPITHELIAL PROTEIN Q9UHB5 93% 32 928 LOST IN NEOPLASM ALPHA. HTEGH03 839477 2792 WUblastx.64 (Q9UHB5) EPITHELIAL PROTEIN Q9UHB5 82% 678 845 LOST IN NEOPLASM ALPHA. 88% 1 732 HTEGY81 637689 2794 WUblastx.64 (Q9CYN5) 5730405I09RIK Q9CYN5 74% 11 544 PROTEIN. HTEHB49 823145 2796 WUblastx.64 (Q9H2T7) RANBP17. Q9H2T7 65% 126 254 99% 203 1537 HTEHV60 637690 2798 WUblastx.64 (Q9D2H5) 4930486B16RIK Q9D2H5 88% 72 1028 PROTEIN. 33% 153 281 89% 1027 1113 HTEHW80 862068 2799 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 71% 297 578 CLONE HEMBA1006036. HTEID25 737852 2800 WUblastx.64 (Q9Y4V7) DJ1178H5.3 (NOVEL Q9Y4V7 100% 259 384 PROTEIN) (FRAGMENT). HTEIJ23 784268 2801 WUblastx.64 (Q9NX98) CDNA FLJ20363 FIS, Q9NX98 93% 107 1303 CLONE HEP17001. HTEIM62 806472 2802 WUblastx.64 (Q9H3C1) PRO0872. Q9H3C1 81% 62 15 76% 123 61 HTEIV33 603393 2803 WUblastx.64 hypothetical protein pir|T47135|T47135 53% 259 11 DKFZp761L0812.1 - human (fragment) HTEJD61 828178 2807 WUblastx.64 (BAB49394) Ml12208 protein. BAB49394 37% 437 682 27% 21 380 30% 437 682 27% 9 383 24% 18 392 29% 21 386 32% 401 682 30% 138 395 24% 452 682 27% 90 380 30% 395 679 28% 437 682 26% 392 592 30% 12 386 HTEJL16 603409 2810 WUblastx.64 (Q9CPU8) 4921511D23RIK Q9CPU8 71% 453 412 PROTEIN. 34% 1042 722 34% 1027 698 80% 412 257 32% 952 728 64% 1030 596 HTEKD35 604979 2813 WUblastx.64 (Q9D6N1) CARBONIC Q9D6N1 89% 40 594 ANHYDRASE (EC 4.2.1.1) (CARBONATE DEHYDRATASE). HTEKP82 694648 2814 WUblastx.64 (Q9D9W1) 1700027A23RIK Q9D9W1 68% 147 689 PROTEIN. HTEKV69 877673 2815 WUblastx.64 (Q9D400) 4933425K02RIK Q9D400 61% 157 1005 PROTEIN. 66% 1052 1096 35% 52 210 HTFOB75 900824 2818 WUblastx.64 (Q9BX86) HP95. Q9BX86 96% 162 2366 50% 2469 2675 21% 1857 2354 HTGAA35 737945 2819 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 64% 1474 1349 CLONE KAIA0536. 78% 1632 1465 HTGAD74 834464 2820 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 66% 665 621 67% 868 668 HTGAP05 637715 2821 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 65% 871 749 CLONE KAIA0536. 73% 990 868 HTGAR21 838159 2823 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 80% 864 820 FOR MGC: 5149). 67% 1081 878 HTGAS70 827320 2824 WUblastx.64 (AAH00037) DNA fragmentation AAH00037 95% 290 1216 factor, 45 kD, alpha p HTGAT65 688864 2825 WUblastx.64 (Q9P0D8) HSPC098 (FRAGMENT). Q9P0D8 42% 1074 1130 60% 1469 1573 HTGAU17 605125 2826 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 58% 621 692 PROTEIN. 44% 703 876 HTGBK95 834490 2828 WUblastx.64 (Q9GMX5) HYPOTHETICAL 12.9 KDA Q9GMX5 66% 126 55 PROTEIN. 70% 235 116 HTGCC01 598903 2829 WUblastx.64 (Q9H387) PRO2550. Q9H387 73% 1099 962 68% 963 784 HTGCK43 828867 2830 WUblastx.64 (P82914) 28S RIBOSOMAL RT15_HUMAN 91% 862 92 PROTEIN S15, MITOCHONDRIAL PRECURSOR HTGDS43 605094 2831 WUblastx.64 (O60921) HUS1+-LIKE PROTEIN. O60921 97% 706 831 HTGDS92 839478 2832 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 69% 1300 1001 CLONE COL04765. HTGEX34 877490 2833 WUblastx.64 (Q9H387) PRO2550. Q9H387 77% 2277 1966 HTGGM37 827310 2835 WUblastx.64 (Q9NUM6) CDNA FLJ11267 FIS, Q9NUM6 76% 1781 1909 CLONE PLACE1009174. HTGGN22 782853 2836 WUblastx.64 (Q9H5R3) CDNA: FLJ23147 FIS, Q9H5R3 57% 741 664 CLONE LNG09295. 66% 682 524 HTHBC58 839916 2838 WUblastx.64 (Q9H387) PRO2550. Q9H387 80% 1267 1208 71% 1452 1294 HTHBQ29 561547 2840 WUblastx.64 (Q9BGV8) HYPOTHETICAL 10.0 KDA Q9BGV8 78% 786 1010 PROTEIN. HTHBZ91 637697 2842 WUblastx.64 (O60448) NEURONAL THREAD O60448 53% 823 668 PROTEIN AD7C-NTP. 61% 948 802 69% 743 705 44% 458 273 52% 947 747 59% 594 466 61% 963 748 50% 360 313 36% 891 802 39% 948 880 57% 569 324 50% 493 425 25% 848 561 48% 408 289 26% 425 249 34% 454 272 30% 513 301 52% 588 367 HTHCA30 637124 2843 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 67% 627 466 FOR MGC: 5149). HTHDB20 669032 2845 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 79% 426 355 CLONE COL04765. 60% 629 429 HTHDF45 621313 2846 WUblastx.64 (Q9H387) PRO2550. Q9H387 42% 987 691 HTHDF86 815686 2847 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 63% 1184 1249 CLONE KAIA0536. 83% 1029 1193 50% 1495 1247 HTHDP65 637699 2849 WUblastx.64 (Q9H387) PRO2550. Q9H387 100% 42 16 66% 205 35 HTHDV50 789402 2851 WUblastx.64 (Q9GMI7) HYPOTHETICAL 9.0 KDA Q9GMI7 47% 299 78 PROTEIN. HTJMA64 775181 2852 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 75% 1065 1211 CLONE COL04765. 75% 1369 1500 52% 1209 1391 44% 1187 1267 64% 893 1087 HTLAD74 638978 2854 WUblastx.64 (P78525) MYB PROTO-ONCOGENE P78525 48% 929 633 PROTEIN (C-MYB). HTLAF81 855970 2855 WUblastx.64 (Q9BUS3) UNKNOWN (PROTEIN Q9BUS3 72% 306 1073 FOR IMAGE: 3461487) (FRAGMENT). HTLBF46 839774 2856 HMMER PFAM: Papain family cysteine PF00112 255 176 751 2.1.1 protease WUblastx.64 (Q9UBX1) CATHEPSIN F CATF_HUMAN 99% 56 757 PRECURSOR (EC 3.4.22.41) (CATSF). HTLBF63 762847 2857 HMMER PFAM: MYND finger PF01753 53.8 652 762 2.1.1 WUblastx.64 (O75800) BLU PROTEIN. O75800 94% 10 792 HTLCX82 847091 2858 WUblastx.64 (Q9BTZ4) SIMILAR TO Q9BTZ4 100% 127 258 EXPRESSED SEQUENCE 2 72% 237 629 EMBRYONIC LETHAL (FRAGMENT). HTLDN34 866199 2860 WUblastx.64 (O94993) SOX30 PROTEIN. O94993 82% 242 751 89% 726 842 69% 9 137 HTLDP19 885353 2861 WUblastx.64 (Q9D5P4) 4930403J07RIK PROTEIN. Q9D5P4 62% 530 751 84% 8 121 61% 159 581 HTLEJ24 608317 2863 WUblastx.64 (Q9D7G6) 2310009N05RIK Q9D7G6 84% 2 619 PROTEIN. HTLEJ75 815631 2864 WUblastx.64 (AAK52668) MMS19. AAK52668 92% 31 2133 HTLEJ75 762849 2865 WUblastx.64 (BAB55315) CDNA FLJ14804 fis, BAB55315 85% 7 798 clone NT2RP4001638, w HTLEP55 637704 2866 WUblastx.64 (AAL37611) Carboxypeptidase A5. AAL37611 97% 1242 1352 92% 1171 1251 53% 561 656 100% 178 561 87% 608 1168 HTLEV80 866200 2867 WUblastx.64 (O15020) BETA-SPECTRIN III O15020 100% 9 452 (FNTA III SPECTRIN). HTLEZ57 634874 2868 WUblastx.64 (Q9P195) PRO1722. Q9P195 57% 662 531 59% 121 26 62% 481 401 HTLFA90 740770 2869 WUblastx.64 (Q9BRY0) UNKNOWN (PROTEIN Q9BRY0 100% 2 1072 FOR IMAGE: 2966557) (FRAGMENT). HTLGL33 835020 2870 WUblastx.64 N-type calcium channel alpha-1 chain, 1 pir|T45115|T45115 22% 364 1092 HTLGQ25 898114 2871 HMMER PFAM: Immunoglobulin domain PF00047 26.6 153 377 2.1.1 WUblastx.64 (Q9H106) DJ576H24.4 (NOVEL Q9H106 100% 114 443 PROTEIN MEMBER OF THE PTPNS (PROTEIN TYROS HTLGS72 897278 2872 WUblastx.64 (Q9JJC0) BRAIN CDNA, CLONE Q9JJC0 34% 6 563 MNCB-2717. HTLGY50 839479 2873 WUblastx.64 (O73884) PUTATIVE O73884 39% 987 1100 PHOSPHATASE. 65% 589 1002 HTLHN86 896930 2874 WUblastx.64 (BAB55144) CDNA FLJ14576 fis, BAB55144 95% 318 938 clone NT2RM4001092, w HTLHN86 838287 2875 WUblastx.64 (BAB55144) CDNA FLJ14576 fis, BAB55144 95% 318 938 clone NT2RM4001092, w HTLHN86 843766 2876 WUblastx.64 (BAB55144) CDNA FLJ14576 fis, BAB55144 95% 318 938 clone NT2RM4001092, w HTLHN86 883351 2877 WUblastx.64 (BAB55144) CDNA FLJ14576 fis, BAB55144 95% 318 938 clone NT2RM4001092, w HTLIW29 899417 2878 HMMER PFAM: Trypsin PF00089 226.4 133 852 2.1.1 WUblastx.64 (Q9Y6M0) TESTISIN PRECURSOR TEST_HUMAN 100% 67 885 (EC 3.4.21.—) (EOSINOPHIL SERIN HTLJC15 898235 2879 WUblastx.64 (Q9D123) 1110032O16RIK Q9D123 71% 996 1715 PROTEIN. HTNAL14 886203 2880 WUblastx.64 (Q9NXU2) CDNA FLJ20054 FIS, Q9NXU2 100% 580 687 CLONE COL00849. HTNBJ15 834872 2882 WUblastx.64 (Q9H055) HYPOTHETICAL 13.8 KDA Q9H055 99% 1536 1889 PROTEIN. HTNBJ15 845783 2883 WUblastx.64 (Q9H055) HYPOTHETICAL 13.8 KDA Q9H055 99% 1536 1889 PROTEIN. HTNBJ15 853916 2884 WUblastx.64 (Q9H055) HYPOTHETICAL 13.8 KDA Q9H055 99% 1536 1889 PROTEIN. HTNBJ15 884024 2885 WUblastx.64 (Q9H055) HYPOTHETICAL 13.8 KDA Q9H055 99% 1536 1889 PROTEIN. HTOAO58 855972 2889 WUblastx.64 (Q9GZW5) SCAN DOMAIN- Q9GZW5 49% 974 1321 CONTAINING PROTEIN 2 (SCAND2). HTOAT56 702026 2890 WUblastx.64 (AAH00407) Synaptogyrin 2. AAH00407 82% 425 673 83% 10 495 HTOBG07 566862 2891 WUblastx.64 (Q9H288) SEROLOGICALLY Q9H288 100% 225 1343 DEFINED BREAST CANCER 41% 498 1343 ANTIGEN NY-BR-16. 37% 438 1343 37% 276 1187 33% 258 1319 34% 462 1304 30% 462 1286 50% 798 953 44% 1182 1343 66% 941 967 25% 261 839 39% 1098 1247 100% 42 71 HTOBG62 566830 2892 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 66% 1378 1112 CLONE COL04765. HTODO45 823125 2895 WUblastx.64 (AAH08373) Similar to hypothetical AAH08373 67% 1408 1226 protein PRO1722. HTOET03 845230 2899 WUblastx.64 (Q9BQC3) SIMILAR TO Q9BQC3 83% 4 1041 DIPTHERIA TOXIN RESISTANCE PROTEIN REQUIRED FOR D HTOET03 837213 2900 WUblastx.64 (Q9BQC3) SIMILAR TO Q9BQC3 83% 4 1041 DIPTHERIA TOXIN RESISTANCE PROTEIN REQUIRED FOR D HTOFA11 637719 2902 WUblastx.64 (Q30086) MHC CLASS II HLA-DQ- Q30086 96% 1348 1635 ALPHA (DR2-DQW1/DR4 DQW3) 80% 1962 2069 (FRAGMENT). 97% 694 942 68% 1882 1968 HTOFC33 824604 2903 WUblastx.64 (Q9H387) PRO2550. Q9H387 65% 900 754 73% 1069 878 HTOGB79 762835 2904 WUblastx.64 (O60448) NEURONAL THREAD O60448 52% 1639 1917 PROTEIN AD7C-NTP. 60% 1642 1806 50% 1661 1825 31% 1465 1530 26% 1238 1528 27% 69 227 60% 1854 1928 43% 1797 1865 32% 1420 1542 75% 2727 2596 52% 2606 2472 42% 1214 1137 77% 2727 2551 49% 2661 2500 42% 2663 2553 56% 1213 1166 45% 2715 2596 68% 1284 1219 41% 1302 1141 55% 1332 1273 48% 1332 1264 52% 2730 2674 55% 1333 1274 55% 1208 1155 60% 1339 1157 62% 2756 2478 HTOHE22 821698 2905 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 75% 1008 1217 CLONE KAIA0536. HTOHG63 834832 2906 WUblastx.64 (Q9H8L6) CDNA FLJ13465 FIS, Q9H8L6 98% 2 670 CLONE PLACE1003493, WEAKLY SIMILAR TO END HTOHJ93 762836 2907 WUblastx.64 (Q9H743) CDNA: FLJ21394 FIS, Q9H743 70% 1323 1048 CLONE COL03536. HTOHN40 843372 2910 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 64% 1889 1839 CLONE KAIA0536. 50% 1845 1654 HTOHR59 762850 2911 WUblastx.64 (AAH07609) Similar to hypothetical AAH07609 92% 1249 1127 protein PRO1722. HTOHS29 722256 2912 WUblastx.64 (AAH08373) Similar to hypothetical AAH08373 80% 129 10 protein PRO1722. HTOID65 636069 2913 WUblastx.64 (Q9H387) PRO2550. Q9H387 56% 630 761 50% 467 658 77% 1420 1316 78% 1314 1165 HTOIE17 688061 2914 WUblastx.64 (Q9NWI4) CDNA FLJ20837 FIS, Q9NWI4 66% 942 889 CLONE ADKA02602. 61% 1078 938 HTOIG16 845999 2915 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 93% 1558 1511 CLONE COL04765. 63% 1793 1563 HTOIH39 839245 2916 WUblastx.64 (O60448) NEURONAL THREAD O60448 53% 750 1106 PROTEIN AD7C-NTP. 46% 53 130 50% 1095 1184 51% 840 1049 57% 64 126 56% 14 121 54% 987 1151 54% 29 133 40% 1039 1167 68% 19 93 HTOJB02 600376 2918 WUblastx.64 (O60448) NEURONAL THREAD O60448 54% 780 652 PROTEIN AD7C-NTP. 33% 1322 1143 50% 1275 1108 66% 919 770 47% 682 626 52% 1235 1161 40% 889 770 50% 787 632 58% 816 727 41% 1424 1338 62% 708 661 37% 784 614 56% 918 658 58% 1411 1265 56% 1274 1134 30% 1275 1093 57% 1403 1251 59% 1424 1098 60% 949 713 HTOJJ26 821702 2919 WUblastx.64 (Q9UHT1) PRO1902 PROTEIN. Q9UHT1 71% 2969 2844 HTOJP25 853623 2920 WUblastx.64 hypothetical L1 protein (third intron of pir|JU0033|JU0033 60% 1388 1519 gene TS) - human HTOJS23 737735 2921 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 57% 1564 1523 CLONE COL04765. 46% 1670 1575 59% 1833 1657 HTOJY56 664504 2922 WUblastx.64 (Q9P1F2) PRO2032. Q9P1F2 100% 88 240 HTPCO75 853645 2925 WUblastx.64 (O00549) ORF2-LIKE PROTEIN O00549 43% 325 26 (FRAGMENT). 36% 1318 1253 HTPDD68 590530 2927 WUblastx.64 (Q9H0G2) HYPOTHETICAL 136.4 KDA Q9H0G2 88% 10 2004 PROTEIN. HTSET62 833692 2930 WUblastx.64 (Q9HCS4) HMG-BOX Q9HCS4 95% 59 124 TRANSCRIPTION FACTOR TCF-3. 66% 124 327 HTSFV18 609939 2931 HMMER PFAM: Low-density lipoprotein PF00057 49.4 170 280 2.1.1 receptor domain class A WUblastx.64 LDL receptor related protein 105 - pir|T00204|T00204 82% 10 984 human HTSGO13 789723 2932 WUblastx.64 (Q9NWJ5) CDNA FLJ20807 FIS, Q9NWJ5 99% 195 548 CLONE ADSE01784. 97% 489 845 HTSGO88 634834 2933 WUblastx.64 (O60448) NEURONAL THREAD O60448 76% 797 612 PROTEIN AD7C-NTP. 52% 640 521 66% 636 574 45% 560 492 69% 791 627 61% 137 75 HTTAH05 772560 2934 WUblastx.64 (Q9H397) PRO2852. Q9H397 60% 548 676 39% 684 926 HTTBJ38 863131 2936 WUblastx.64 (Q9VST7) CG4911 PROTEIN. Q9VST7 25% 497 1177 HTTDB11 638132 2937 WUblastx.64 (Q99LX8) UNKNOWN (PROTEIN Q99LX8 100% 130 240 FOR MGC: 7346). HTTDG27 608318 2938 WUblastx.64 (O62658) LINE-1 ELEMENT ORF2. O62658 44% 454 380 35% 392 3 HTTDN24 766485 2939 WUblastx.64 (Q9BVN5) HYPOTHETICAL 120.6 KDA Q9BVN5 95% 628 1725 PROTEIN. 32% 937 1593 95% 3 629 32% 1114 1596 HTTDO33 899418 2940 WUblastx.64 (Q96GQ9) Unknown (protein for Q96GQ9 95% 22 753 MGC: 16648). HTTEO25 853403 2942 WUblastx.64 (Q9BWK9) UNKNOWN (PROTEIN Q9BWK9 97% 1314 1424 FOR IMAGE: 2900813) (FRAGMENT). HTTEP11 562023 2943 WUblastx.64 hypothetical protein pir|T46441|T46441 70% 753 830 DKFZp434C0927.1 - human 74% 1006 1110 51% 812 928 100% 625 753 HTTES77 844417 2944 WUblastx.64 (AAK40083) Inhibin binding protein AAK40083 82% 697 2085 long isoform. 37% 3 725 31% 6 713 30% 93 704 36% 659 733 39% 697 1818 34% 697 1815 33% 982 1818 88% 3 719 33% 697 1827 31% 1096 1821 31% 736 1833 35% 45 719 28% 901 1761 35% 6 737 34% 9 680 32% 45 716 36% 1240 1818 HTTFG15 600377 2946 WUblastx.64 (Q9GMK2) HYPOTHETICAL 10.0 KDA Q9GMK2 66% 327 482 PROTEIN. HTWAM19 618318 2947 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 76% 1852 1763 CLONE COL04765. 64% 1764 1525 HTWBO30 655371 2949 WUblastx.64 (Q9N083) UNNAMED PORTEIN Q9N083 47% 560 453 PRODUCT. 55% 751 578 HTWBZ57 828030 2950 WUblastx.64 (O95273) D-TYPE CYCLIN- O95273 87% 218 1276 INTERACTING PROTEIN 1. HTWCC10 747687 2951 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 60% 1051 947 CLONE KAIA0536. 63% 1241 1095 HTWCE14 762839 2952 WUblastx.64 (Q9BSZ7) UNKNOWN (PROTEIN Q9BSZ7 100% 121 231 FOR MGC: 4322). HTWCT76 767758 2953 WUblastx.64 (Q9BGW3) HYPOTHETICAL 13.5 KDA Q9BGW3 68% 1550 1368 PROTEIN. 58% 1361 1311 HTWDJ17 784037 2954 WUblastx.64 (Q9CYV6) 2810439F02RIK Q9CYV6 93% 17 1255 PROTEIN. HTWDM89 569787 2955 WUblastx.64 (Q9P195) PRO1722. Q9P195 63% 1413 1267 61% 1277 1098 HTWEQ36 823369 2958 WUblastx.64 (Q9NXK9) CDNA FLJ20187 FIS, Q9NXK9 68% 748 891 CLONE COLF0433. HTWFA88 596942 2960 WUblastx.64 probable transposase - human pir|S72481|S72481 58% 1286 1357 transposon MER37 70% 1350 1559 51% 709 1272 HTWFO43 858741 2962 WUblastx.64 (O60448) NEURONAL THREAD O60448 67% 890 627 PROTEIN AD7C-NTP. 62% 1241 1056 63% 754 689 62% 1108 980 65% 1242 1102 61% 905 744 53% 684 601 40% 790 665 58% 675 625 34% 1184 975 35% 863 744 61% 1257 1039 45% 1063 992 64% 657 607 40% 1215 1087 34% 778 659 100% 1016 996 HTXAD75 745410 2965 WUblastx.64 (Q9GLG1) CALPAIN 2. Q9GLG1 92% 345 917 HTXAR92 656935 2966 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 77% 1578 1297 CLONE COL04765. HTXBU88 604985 2968 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 75% 771 736 CLONE KAIA0536. 65% 739 680 60% 701 453 HTXCP27 892364 2969 WUblastx.64 (AAH00057) Protein phosphatase 1G AAH00057 97% 521 628 (formerly 2C), ma 100% 946 1149 98% 285 524 HTXCU30 839846 2970 WUblastx.64 (Q9H728) CDNA: FLJ21463 FIS, Q9H728 81% 706 641 CLONE COL04765. 80% 639 577 59% 875 684 HTXCV44 740773 2971 WUblastx.64 (Q9HA67) CDNA FLJ12155 FIS, Q9HA67 52% 883 945 CLONE MAMMA1000472. 70% 809 898 63% 630 803 63% 1521 1366 68% 1370 1323 HTXDJ75 562780 2973 WUblastx.64 (Q26195) PVA1 GENE. Q26195 51% 1598 1335 HTXDT72 778083 2977 WUblastx.64 (Q9BSV9) SIMILAR TO 95 KDA Q9BSV9 100% 8 400 RETINOBLASTOMA PROTEIN BINDING PROTEIN, KI HTXDZ68 824545 2979 WUblastx.64 (Q9P0D0) HSPC106 (FRAGMENT). Q9P0D0 72% 80 277 54% 136 498 HTXEN33 589556 2980 WUblastx.64 (Q99419) ICSAT TRANSCRIPTION Q99419 78% 843 968 FACTOR (FRAGMENT). HTXJD08 566885 2985 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 59% 743 1111 PROTEIN (FRAGMENT). 47% 604 756 45% 395 583 HTXJD85 840391 2986 WUblastx.64 (Q9HAD8) CDNA FLJ11786 FIS, Q9HAD8 52% 1093 818 CLONE HEMBA1006036. HTXJI59 869905 2988 WUblastx.64 (O15290) P66SHC. O15290 93% 1708 1571 HTXJJ92 688062 2989 WUblastx.64 (Q9N044) UNNAMED PROTEIN Q9N044 59% 709 614 PRODUCT. 66% 611 531 HTXJM94 853413 2990 WUblastx.64 (Q9Y2S9) HSPC019. Q9Y2S9 83% 44 262 HTXJW06 789201 2992 WUblastx.64 (Q9UI28) ADRENAL GLAND Q9UI28 100% 868 623 PROTEIN AD-003. HTXKB57 844329 2993 WUblastx.64 (Q9P073) HSPC309 (FRAGMENT). Q9P073 94% 1072 1128 65% 1125 1523 HTXKH40 600378 2995 WUblastx.64 (Q9Y380) CGI-71 PROTEIN. Q9Y380 94% 9 290 HTXKK76 858755 2996 WUblastx.64 pro-pol-dUTPase polyprotein - murine pir|T29097|T29097 44% 466 2 endogenous retrovirus ERV-L (fragment) HTXKL53 783141 2997 WUblastx.64 (Q9Y3F0) CGI-05 PROTEIN. Q9Y3F0 100% 1315 1359 90% 3 89 HTXLC05 843523 3000 WUblastx.64 (Q9H189) SPHINGOSINE-1- Q9H189 25% 4 531 PHOSPHATASE. HTXLY94 844165 3003 WUblastx.64 hypothetical protein pir|T14744|T14744 64% 77 643 DKFZp586F0424.1 - human (fragment) HTXNV66 840597 3004 WUblastx.64 (Q9UJY4) ADP-RIBOSYLATION GGA2_HUMAN 99% 18 431 FACTOR BINDING PROTEIN GGA2 (GOLG HTXOW27 839280 3006 WUblastx.64 (Q9P2V7) PROTEIN CONTAINING Q9P2V7 100% 1652 2086 CXXC DOMAIN 1 92% 1309 1656 (HYPOTHETICAL 75.7 KDA PROT 78% 229 1188 HTXPD86 834772 3007 WUblastx.64 (Q9UGP9) WD-REPEAT PROTEIN 5 WDR5_HUMAN 95% 74 1132 (FRAGMENT). HTXPT57 897834 3008 WUblastx.64 (O60448) NEURONAL THREAD O60448 60% 939 838 PROTEIN AD7C-NTP. 52% 693 544 42% 564 451 53% 471 427 26% 722 543 35% 725 474 37% 1123 938 45% 974 849 53% 705 526 40% 1054 911 53% 911 834 75% 1104 1069 64% 891 841 51% 551 432 57% 425 384 52% 1084 926 HTYSJ88 598718 3009 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 64% 729 457 CLONE KAIA0536. HUFAB57 587275 3012 WUblastx.64 ISOFORM B OF Q9UNP9 sp_vs|Q9UNP9- 94% 1266 1316 01|Q9UNP9 53% 826 1035 HUFAO92 608175 3014 WUblastx.64 (Q9D922) 1810010G06RIK Q9D922 69% 232 522 PROTEIN. HUFAO94 668267 3015 WUblastx.64 (Q9UI14) PRENYLATED RAB Q9UI14 100% 3 551 ACCEPTOR 1. HUFAP33 600379 3016 HMMER PFAM: Zona pellucida-like domain PF00100 223.2 282 1016 2.1.1 WUblastx.64 (Q9Y211) DMBT1 PROTEIN. Q9Y211 93% 228 1016 100% 26 226 43% 29 217 41% 445 546 38% 445 546 38% 445 522 100% 1013 1117 HUFBV62 742892 3020 WUblastx.64 (Q9CWU4) 2410004B18RIK Q9CWU4 76% 76 264 PROTEIN. HUKAD46 604986 3024 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 46% 589 747 CLONE KAIA0536. 63% 377 598 HUKCS86 826468 3027 WUblastx.64 (Q9NX09) CDNA FLJ20500 FIS, Q9NX09 82% 201 896 CLONE KAT09159. HUKCS86 844245 3028 WUblastx.64 (Q9NX09) CDNA FLJ20500 FIS, Q9NX09 82% 201 896 CLONE KAT09159. HUKEA22 702029 3029 WUblastx.64 (Q9PTD7) CINGULIN. Q9PTD7 68% 575 12 HUKFC71 1300740 3032 WUblastx.64 (P82914) 28S RIBOSOMAL RT15_HUMAN 97% 292 414 PROTEIN S15, MITOCHONDRIAL 65% 408 929 PRECURSOR HUKFC71 845161 3111 blastx.2 MITOCHONDRIAL 28S sp|P82914|P82914 100% 293 931 RIBOSOMAL PROTEIN S15 (MRP- S15). HUKFV37 844644 3033 WUblastx.64 (AAH00407) Synaptogyrin 2. AAH00407 91% 49 720 HUNAL39 559448 3035 WUblastx.64 (Q14288) HYPOTHETICAL Q14288 81% 1009 629 PROTEIN (FRAGMENT). HUSAO04 877889 3036 HMMER PFAM: tRNA synthetases class I (W PF00579 260.4 255 965 2.1.1 and Y) WUblastx.64 (Q9H817) CDNA FLJ13995 FIS, Q9H817 92% 30 1451 CLONE Y79AA1002209, WEAKLY SIMILAR TO TYR HUSAO04 870791 3037 HMMER PFAM: tRNA synthetases class I (W PF00579 212.2 297 965 2.1.1 and Y) WUblastx.64 (Q9H817) CDNA FLJ13995 FIS, Q9H817 92% 30 1451 CLONE Y79AA1002209, WEAKLY SIMILAR TO TYR HUSCA09 853417 3038 HMMER PFAM: Leucine Rich Repeat PF00560 69.9 1259 1327 2.1.1 WUblastx.64 (Q9H075) HYPOTHETICAL 81.8 KDA Q9H075 52% 44 1615 PROTEIN. HUSCJ01 740775 3039 WUblastx.64 (Q9CWP6) 2410013I23RIK Q9CWP6 70% 1 252 PROTEIN. HUSGB23 704094 3040 WUblastx.64 (Q9H0F5) HYPOTHETICAL 43.9 KDA Q9H0F5 97% 143 247 PROTEIN. HUSGY15 759888 3043 WUblastx.64 (Q9JKN1) ZINC TRANSPORTER Q9JKN1 76% 162 1289 LIKE 2 (1810059J10RIK PROTEIN). HUSHD41 866498 3044 WUblastx.64 (BAB55441) CDNA FLJ14993 fis, BAB55441 86% 2575 1541 clone Y79AA1001874, w 50% 2569 2483 HUSHK65 847188 3045 HMMER PFAM: CUB domain PF00431 292.1 887 1222 2.1.1 WUblastx.64 (AAG41897) Neuropilin-2a(17). AAG41897 97% 806 2122 95% 2727 3023 82% 3046 3579 92% 2089 2721 32% 2086 2619 31% 1547 2086 92% 3008 3046 HUSIK45 853932 3046 WUblastx.64 (Q96CN8) Hypothetical 30.9 kDa Q96CN8 91% 23 454 protein (Fragment). HUSIO57 886206 3047 WUblastx.64 (Q9Y3E2) HYPOTHETICAL YCE3_HUMAN 100% 170 565 PROTEIN CGI-143. HUSIR70 838163 3049 WUblastx.64 (Q9HDC9) BSCV PROTEIN Q9HDC9 97% 1 345 (FRAGMENT). HUSXP50 561962 3050 WUblastx.64 (Q63777) HYPOTHETICAL 32.0 KDA Q63777 26% 178 291 PROTEIN. 57% 27 68 55% 113 172 HUVCQ68 561963 3053 WUblastx.64 (Q9BV40) SIMILAR TO VESICLE- Q9BV40 82% 40 339 ASSOCIATED MEMBRANE PROTEIN 8 (ENDOBREVIN HUVEG53 797556 3055 WUblastx.64 (O62658) LINE-1 ELEMENT ORF2. O62658 35% 1843 1676 39% 2148 1939 60% 1391 1263 HWAAH11 815548 3056 WUblastx.64 (Q9HAB3) CDNA FLJ11856 FIS, Q9HAB3 74% 362 754 CLONE HEMBA1006789 (SIMILAR TO HYPOTHETIC HWAAQ28 806588 3057 WUblastx.64 (Q9NZZ8) HSPC169 Q9NZZ8 96% 62 676 (HYPOTHETICAL 33.9 KDA 23% 649 801 PROTEIN). 100% 651 980 HWAAY60 745917 3058 WUblastx.64 (Q9BQA1) BA552M11.2.2 (NOVEL Q9BQA1 76% 504 722 PROTEIN (ISOFORM 2)) (UNKNOWN) (PROTEIN HWABR43 823367 3059 WUblastx.64 (Q9BVD9) UNKNOWN (PROTEIN Q9BVD9 75% 2081 2034 FOR MGC: 5149). 79% 2266 2093 HWACZ33 827311 3061 WUblastx.64 (Q9CTA6) 1110035E02RIK Q9CTA6 62% 389 436 PROTEIN (FRAGMENT). 73% 234 392 HWADV90 897732 3062 WUblastx.64 (Q9NX17) CDNA FLJ20489 FIS, Q9NX17 78% 2375 2163 CLONE KAT08285. HWAEB52 873207 3063 WUblastx.64 (Q9H8U5) CDNA FLJ13219 FIS, Q9H8U5 68% 878 1426 CLONE NT2RP4001849, WEAKLY 60% 389 685 SIMILAR TO SH3 23% 356 667 28% 874 1113 31% 1191 1286 40% 344 442 35% 299 466 HWBAK71 853581 3064 WUblastx.64 (Q9T9V8) NADH Q9T9V8 87% 77 175 DEHYDROGENASE SUBUNIT 3. 83% 305 340 HWBBU75 780360 3065 WUblastx.64 (Q9R189) MUNC13-4 PROTEIN. Q9R189 82% 1454 2362 73% 913 1434 80% 194 952 62% 2229 2729 31% 1586 1711 34% 401 532 HWBCN81 853580 3066 WUblastx.64 (Q9H101) DJ776F14.2 (A NOVEL Q9H101 94% 195 560 PROTEIN MEMBER OF THE PTPNS 33% 222 560 (PROTEIN TYR 100% 150 221 HWBCX93 853418 3068 WUblastx.64 (Q9NX85) CDNA FLJ20378 FIS, Q9NX85 80% 642 553 CLONE KAIA0536. 84% 796 641 HWHGV77 788555 3072 WUblastx.64 (Q9D6W1) 2310050C09RIK Q9D6W1 70% 136 621 PROTEIN. 51% 127 552 37% 127 588 47% 124 318 100% 721 756 HWHGW09 702032 3073 HMMER PFAM: Uncharacterized protein family PF01027 68.3 271 444 2.1.1 WUblastx.64 (Q9Y3C2) CGI-119 PROTEIN. Q9Y3C2 84% 256 450 HWHPU44 778090 3075 WUblastx.64 (Q9H1C8) PUTATIVE Q9H1C8 97% 311 547 CYTOPLASMATIC PROTEIN. HWLAT50 840753 3077 WUblastx.64 (Q99JR4) UNKNOWN (PROTEIN Q99JR4 48% 1864 2106 FOR IMAGE: 3599558) (FRAGMENT). HWLGP26 834770 3079 WUblastx.64 (Q9NP87) DNA POLYMERASE MU. Q9NP87 93% 674 760 100% 269 298 94% 295 465 87% 432 623 100% 3 254 HWLHO31 837480 3080 WUblastx.64 (Q9NQT8) KINESIN-LIKE PROTEIN Q9NQT8 100% 41 205 GAKIN. 96% 190 1674 HWLJN08 800626 3082 WUblastx.64 (Q9BX68) HIT-17 KDA. Q9BX68 99% 434 844 HWLRE03 831198 3083 WUblastx.64 (O60646) HYPOTHETICAL 53.8 KDA O60646 100% 1008 1232 PROTEIN (FRAGMENT). 99% 1238 2431 HWTBL86 791720 3089 WUblastx.64 hypothetical protein pir|T17285|T17285 91% 1 1080 DKFZp434N0535.1 - human (fragment) HWTBX66 732187 3090 WUblastx.64 (Q9BRI8) UNKNOWN (PROTEIN Q9BRI8 71% 63 1073 FOR MGC: 11332). HYAAD61 897736 3092 WUblastx.64 (O21101) ATPASE 6/8 O21101 56% 2009 2179 (FRAGMENT). HYBAP75 853561 3094 WUblastx.64 (Q9N032) UNNAMED PROTEIN Q9N032 64% 909 1025 PRODUCT. RACE Protocol for Recovery of Full-Length Genes

Partial cDNA clones can be made full-length by utilizing the rapid amplification of cDNA ends (RACE) procedure described in Frohman, M. A., et al., Proc. Nat'l. Acad. Sci. USA, 85:8998-9002 (1988). A cDNA clone missing either the 5′ or 3′ end can be reconstructed to include the absent base pairs extending to the translational start or stop codon, respectively. In some cases, cDNAs are missing the start codon of translation, therefor. The following briefly describes a modification of this original 5′ RACE procedure. Poly A+ or total RNA is reverse transcribed with Superscript II (Gibco/BRL) and an antisense or complementary primer specific to the cDNA sequence. The primer is removed from the reaction with a Microcon Concentrator (Amicon). The first-strand cDNA is then tailed with dATP and terminal deoxynucleotide transferase (Gibco/BRL). Thus, an anchor sequence is produced which is needed for PCR amplification. The second strand is synthesized from the dA-tail in PCR buffer, Taq DNA polymerase (Perkin-Elmer Cetus), an oligo-dT primer containing three adjacent restriction sites (XhoI, SalI and ClaI) at the 5′ end and a primer containing just these restriction sites. This double-stranded cDNA is PCR amplified for 40 cycles with the same primers as well as a nested cDNA-specific antisense primer. The PCR products are size-separated on an ethidium bromide-agarose gel and the region of gel containing cDNA products the predicted size of missing protein-coding DNA is removed. cDNA is purified from the agarose with the Magic PCR Prep kit (Promega), restriction digested with XhoI or SalI, and ligated to a plasmid such as pBluescript SKII (Stratagene) at XhoI and EcoRV sites. This DNA is transformed into bacteria and the plasmid clones sequenced to identify the correct protein-coding inserts. Correct 5′ ends are confirmed by comparing this sequence with the putatively identified homologue and overlap with the partial cDNA clone. Similar methods known in the art and/or commercial kits are used to amplify and recover 3′ ends.

Several quality-controlled kits are commercially available for purchase. Similar reagents and methods to those above are supplied in kit form from Gibco/BRL for both 5′ and 3′ RACE for recovery of full length genes. A second kit is available from Clontech which is a modification of a related technique, SLIC (single-stranded ligation to single-stranded cDNA), developed by Dumas et al., Nucleic Acids Res., 19:5227-32 (1991). The major differences in procedure are that the RNA is alkaline hydrolyzed after reverse transcription and RNA ligase is used to join a restriction site-containing anchor primer to the first-strand cDNA. This obviates the necessity for the dA-tailing reaction which results in a polyT stretch that is difficult to sequence past.

An alternative to generating 5′ or 3′ cDNA from RNA is to use cDNA library double-stranded DNA. An asymmetric PCR-amplified antisense cDNA strand is synthesized with an antisense cDNA-specific primer and a plasmid-anchored primer. These primers are removed and a symmetric PCR reaction is performed with a nested cDNA-specific antisense primer and the plasmid-anchored primer.

RNA Ligase Protocol for Generating the 5′ or 3′ End Sequences to Obtain Full Length Genes

Once a gene of interest is identified, several methods are available for the identification of the 5′ or 3′ portions of the gene which may not be present in the original cDNA plasmid. These methods include, but are not limited to, filter probing, clone enrichment using specific probes and protocols similar and identical to 5′ and 3′ RACE. While the full length gene may be present in the library and can be identified by probing, a useful method for generating the 5′ or 3′ end is to use the existing sequence information from the original cDNA to generate the missing information. A method similar to 5′ RACE is available for generating the missing 5′ end of a desired full-length gene. (This method was published by Fromont-Racine et al., Nucleic Acids Res., 21(7):1683-1684 (1993)). Briefly, a specific RNA oligonucleotide is ligated to the 5′ ends of a population of RNA presumably containing full-length gene RNA transcript and a primer set containing a primer specific to the ligated RNA oligonucleotide and a primer specific to a known sequence of the gene of interest, is used to PCR amplify the 5′ portion of the desired full length gene which may then be sequenced and used to generate the full length gene. This method starts with total RNA isolated from the desired source, poly A RNA may be used but is not a prerequisite for this procedure. The RNA preparation may then be treated with phosphatase if necessary to eliminate 5′ phosphate groups on degraded or damaged RNA which may interfere with the later RNA ligase step. The phosphatase if used is then inactivated and the RNA is treated with tobacco acid pyrophosphatase in order to remove the cap structure present at the 5′ ends of messenger RNAs. This reaction leaves a 5′ phosphate group at the 5′ end of the cap cleaved RNA which can then be ligated to an RNA oligonucleotide using T4 RNA ligase. This modified RNA preparation can then be used as a template for first strand cDNA synthesis using a gene specific oligonucleotide. The first strand synthesis reaction can then be used as a template for PCR amplification of the desired 5′ end using a primer specific to the ligated RNA oligonucleotide and a primer specific to the known sequence of the gene of interest. The resultant product is then sequenced and analyzed to confirm that the 5′ end sequence belongs to the relevant gene.

The present invention also relates to vectors or plasmids which include such DNA sequences, as well as the use of the DNA sequences. The material deposited with the ATCC (e.g., as described in columns 2 and 3 of Table 1A, and/or as set forth in Table 1B, Table 6, or Table 7) is a mixture of cDNA clones derived from a variety of human tissue and cloned in either a plasmid vector or a phage vector, as described, for example, in Table 1A and Table 7. These deposits are referred to as “the deposits” herein. The tissues from which some of the clones were derived are listed in Table 7, and the vector in which the corresponding cDNA is contained is also indicated in Table 7. The deposited material includes cDNA clones corresponding to SEQ ID NO:X described, for example, in Table 1A and/or 1B (ATCC Deposit No:Z). A clone which is isolatable from the ATCC Deposits by use of a sequence listed as SEQ ID NO:X, may include the entire coding region of a human gene or in other cases such clone may include a substantial portion of the coding region of a human gene. Furthermore, although the sequence listing may in some instances list only a portion of the DNA sequence in a clone included in the ATCC Deposits, it is well within the ability of one skilled in the art to sequence the DNA included in a clone contained in the ATCC Deposits by use of a sequence (or portion thereof) described in, for example Tables 1A and/or 1B or 2, by procedures hereinafter further described, and others apparent to those skilled in the art.

Also provided in Table 1A and 7 is the name of the vector which contains the cDNA clone. Each vector is routinely used in the art. The following additional information is provided for convenience.

Vectors Lambda Zap (U.S. Pat. Nos. 5,128,256 and 5,286,636), Uni-Zap XR (U.S. Pat. Nos. 5,128,256 and 5,286,636), Zap Express (U.S. Pat. Nos. 5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. et al., Nucleic Acids Res. 16:7583-7600 (1988); Alting-Mees, M. A. and Short, J. M., Nucleic Acids Res. 17:9494 (1989)) and pBK (Alting-Mees, M. A. et al., Strategies 5:58-61 (1992)) are commercially available from Stratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La Jolla, Calif., 92037. pBS contains an ampicillin resistance gene and pBK contains a neomycin resistance gene. Phagemid pBS may be excised from the Lambda Zap and Uni-Zap XR vectors, and phagemid pBK may be excised from the Zap Express vector. Both phagemids may be transformed into E. coli strain XL-1 Blue, also available from Stratagene.

Vectors pSport1, pCMVSport 1.0, pCMVSport 2.0 and pCMVSport 3.0, were obtained from Life Technologies, Inc., P.O. Box 6009, Gaithersburg, Md. 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. See, for instance, Gruber, C. E., ct al., Focus 15:59-(1993). Vector lafmid BA (Bento Soares, Columbia University, New York, N.Y.) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Bluc. Vector pCR®2.1, which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, Calif. 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. See, for instance, Clark, J. M., Nuc. Acids Res. 16:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991).

The present invention also relates to the genes corresponding to SEQ ID NO:X, SEQ ID NO:Y, and/or the deposited clone (ATCC Deposit No:Z). The corresponding gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material.

Also provided in the present invention are allelic variants, orthologs, and/or species homologs. Procedures known in the art can be used to obtain full-length genes, allelic variants, splice variants, full-length coding portions, orthologs, and/or species homologs of genes corresponding to SEQ ID NO:X or the complement thereof, polypeptides encoded by genes corresponding to SEQ ID NO:X or the complement thereof, and/or the cDNA contained in ATCC Deposit No:Z, using information from the sequences disclosed herein or the clones deposited with the ATCC. For example, allelic variants and/or species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for allelic variants and/or the desired homologue.

The polypeptides of the invention can be prepared in any suitable manner. Such polypeptides include isolated naturally occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well understood in the art.

The polypeptides may be in the form of the secreted protein, including the mature form, or may be a part of a larger protein, such as a fusion protein (see below). It is often advantageous to include an additional amino acid sequence which contains secretory or leader sequences, pro-sequences, sequences which aid in purification, such as multiple histidine residues, or an additional sequence for stability during recombinant production.

The polypeptides of the present invention are preferably provided in an isolated form, and preferably are substantially purified. A recombinantly produced version of a polypeptide, including the secreted polypeptide, can be substantially purified using techniques described herein or otherwise known in the art, such as, for example, by the one-step method described in Smith and Johnson, Gene 67:31-40 (1988). Polypeptides of the invention also can be purified from natural, synthetic or recombinant sources using techniques described herein or otherwise known in the art, such as, for example, antibodies of the invention raised against the polypeptides of the present invention in methods which are well known in the art.

The present invention provides a polynucleotide comprising, or alternatively consisting of, the nucleic acid sequence of SEQ ID NO:X, and/or the cDNA sequence contained in ATCC Deposit No:Z. The present invention also provides a polypeptide comprising, or alternatively, consisting of, the polypeptide sequence of SEQ ID NO:Y, a polypeptide encoded by SEQ ID NO:X or a complement thereof, a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or the polypeptide sequence encoded by a nucleotide sequence in SEQ ID NO:B as defined in column 6 of Table 1C. Polynucleotides encoding a polypeptide comprising, or alternatively consisting of the polypeptide sequence of SEQ ID NO:Y, a polypeptide encoded by SEQ ID NO:X, a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or a polypeptide sequence encoded by a nucleotide sequence in SEQ ID NO:B as defined in column 6 of Table 1C are also encompassed by the invention. The present invention further encompasses a polynucleotide comprising, or alternatively consisting of, the complement of the nucleic acid sequence of SEQ ID NO:X, a nucleic acid sequence encoding a polypeptide encoded by the complement of the nucleic acid sequence of SEQ ID NO:X, and/or the cDNA contained in ATCC Deposit No:Z.

Moreover, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in Table 1C column 6, or any combination thereof. Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in Table 1C column 6, or any combination thereof. In further embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table 1C, column 6, and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ID NO:B (see Table 1C, column 5). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table 1C, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table 1C, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.

Further, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1C which correspond to the same Clone ID (see Table 1C, column 1), or any combination thereof. Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in column 6 of Table 1C which correspond to the same Clone ID (see Table 1C, column 1), or any combination thereof. In further embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1C which correspond to the same Clone ID (see Table 1C, column 1) and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ID NO:B (see Table 1C, column 5). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1C which correspond to the same Clone ID (see Table 1C, column 1) and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1C which correspond to the same Clone ID (see Table 1C, column 1) and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.

Further, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1C which correspond to the same contig sequence identifier SEQ ID NO:X (see Table 1C, column 2), or any combination thereof. Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in column 6 of Table 1C which correspond to the same contig sequence identifier SEQ ID NO:X (see Table 1C, column 2), or any combination thereof. In further embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1C which correspond to the same contig sequence identifier SEQ ID NO:X (see Table 1C, column 2) and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ID NO:B (see Table 1C, column 5). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1C which correspond to the same contig sequence identifier SEQ ID NO:X (see Table 1C, column 2) and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1C which correspond to the same contig sequence identifier SEQ ID NO:X (see Table 1C, column 2) and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ID NO:A (See Table 1C, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.

Moreover, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in the same row of Table 1C column 6, or any combination thereof. Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in the same row of Table 1C column 6, or any combination thereof. In preferred embodiments, the polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in the same row of Table 1C column 6, wherein sequentially delineated sequences in the table (i.e. corresponding to those exons located closest to each other) are directly contiguous in a 5′ to 3′ orientation. In further embodiments, above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table 1C, column 6, and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ID NO:B (see Table 1C, column 5). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table 1C, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table 1C, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1C, and the polynucleotide sequence of SEQ ID NO:X (e.g., as defined in Table 1C, column 2) or fragments or variants thereof. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1C which correspond to the same Clone ID (see Table 1C, column 1), and the polynucleotide sequence of SEQ ID NO:X (e.g., as defined in Table 1A, 1B, or 1C) or fragments or variants thereof. In preferred embodiments, the delineated sequence(s) and polynucleotide sequence of SEQ ID NO:X correspond to the same Clone ID. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.

In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in the same row of column 6 of Table 1C, and the polynucleotide sequence of SEQ ID NO:X (e.g., as defined in Table 1A, 1B, or 1C) or fragments or variants thereof. In preferred embodiments, the delineated sequence(s) and polynucleotide sequence of SEQ ID NO:X correspond to the same row of column 6 of Table 1C. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of the sequence of SEQ ID NO:X are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of a fragment or variant of the sequence of SEQ ID NO:X are directly contiguous Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3′ 10 polynucleotides of the sequence of SEQ ID NO:X and the 5′ 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table 1C are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3′ 10 polynucleotides of a fragment or variant of the sequence of SEQ ID NO:X and the 5′ 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table 1C are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides, are also encompassed by the invention.

In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of another sequence in column 6 are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of another sequence in column 6 corresponding to the same Clone ID (see Table 1C, column 1) are directly contiguous. Nucleic acids which hybridize to the complement of these 20 lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3′ 10 polynucleotides of one sequence in column 6 corresponding to the same contig sequence identifier SEQ ID NO:X (see Table 1C, column 2) are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of another sequence in column 6 corresponding to the same row are directly contiguous. In preferred embodiments, the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C is directly contiguous with the 5′ 10 polynucleotides of the next sequential exon delineated in Table 1C, column 6. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

Table 3

Many polynucleotide sequences, such as EST sequences, are publicly available and accessible through sequence databases and may have been publicly available prior to conception of the present invention. Preferably, such related polynucleotides are specifically excluded from the scope of the present invention. Accordingly, for each contig sequence (SEQ ID NO:X) listed in the fifth column of Table 1A and/or the fourth column of Table 1B.1, preferably excluded are one or more polynucleotides comprising a nucleotide sequence described by the general formula of a-b, where a is any integer between 1 and the final nucleotide minus 15 of SEQ ID NO:X, b is an integer of 15 to the final nucleotide of SEQ ID NO:X, where both a and b correspond to the positions of nucleotide residues shown in SEQ ID NO:X, and where b is greater than or equal to a+14. More specifically, preferably excluded are one or more polynucleotides comprising a nucleotide sequence described by the general formula of a-b, where a and b are integers as defined in columns 4 and 5, respectively, of Table 3. In specific embodiments, the polynucleotides of the invention do not consist of at least one, two, three, four, five, ten, or more of the specific polynucleotide sequences referenced by the Genbank Accession No. as disclosed in column 6 of Table 3 (including for example, published sequence in connection with a particular BAC clone). In further embodiments, preferably excluded from the invention are the specific polynucleotide sequence(s) contained in the clones corresponding to at least one, two, three, four, five, ten, or more of the available material having the accession numbers identified in the sixth column of this Table (including for example, the actual sequence contained in an identified BAC clone). In no way is this listing meant to encompass all of the sequences which may be excluded by the general formula, it is just a representative example. All references available through these accessions are hereby incorporated by reference in their entirety. LENGTHY TABLE REFERENCED HERE US20070032413A1-20070208-T00004 Please refer to the end of the specification for access instructions. Description of Table 4

Table 4 provides a key to the tissue/cell source identifier code disclosed in Table 1B.2, column 5. Column 1 of Table 4 provides the tissue/cell source identifier code disclosed in Table 1B.2, Column 5. Columns 2-5 provide a description of the tissue or cell source. Note that “Description” and “Tissue” sources (i.e. columns 2 and 3) having the prefix “a_” indicates organs, tissues, or cells derived from “adult” sources. Codes corresponding to diseased tissues are indicated in column 6 with the word “disease.” The use of the word “disease” in column 6 is non-limiting. The tissue or cell source may be specific (e.g. a neoplasm), or may be disease-associated (e.g., a tissue sample from a normal portion of a diseased organ). Furthermore, tissues and/or cells lacking the “disease” designation may still be derived from sources directly or indirectly involved in a disease state or disorder, and therefore may have a further utility in that disease state or disorder. In numerous cases where the tissue/cell source is a library, column 7 identifies the vector used to generate the library. TABLE 4 Code Description Tissue Organ Cell Line Disease Vector AR022 a_Heart a_Heart AR023 a_Liver a_Liver AR024 a_mammary gland a_mammary gland AR025 a_Prostate a_Prostate AR026 a_small intestine a_small intestine AR027 a_Stomach a_Stomach AR028 Blood B cells Blood B cells AR029 Blood B cells activated Blood B cells activated AR030 Blood B cells resting Blood B cells resting AR031 Blood T cells activated Blood T cells activated AR032 Blood T cells resting Blood T cells resting AR033 brain brain AR034 breast breast AR035 breast cancer breast cancer AR036 Cell Line CAOV3 Cell Line CAOV3 AR037 cell line PA-1 cell line PA-1 AR038 cell line transformed cell line transformed AR039 colon colon AR040 colon (9808co65R) colon (9808co65R) AR041 colon (9809co15) colon (9809co15) AR042 colon cancer colon cancer AR043 colon cancer (9808co64R) colon cancer (9808co64R) AR044 colon cancer 9809co14 colon cancer 9809co14 AR050 Donor II B Cells 24 hrs Donor II B Cells 24 hrs AR051 Donor II B Cells 72 hrs Donor II B Cells 72 hrs AR052 Donor II B-Cells 24 hrs. Donor II B-Cells 24 hrs AR053 Donor II B-Cells 72 hrs Donor II B-Cells 72 hrs AR054 Donor II Resting B Cells Donor II Resting B Cells AR055 Heart Heart AR056 Human Lung (clonetech) Human Lung (clonetech) AR057 Human Mammary (clontech) Human Mammary (clontech) AR058 Human Thymus (clonetech) Human Thymus (clonetech) AR059 Jurkat (unstimulated) Jurkat (unstimulated) AR060 Kidney Kidney AR061 Liver Liver AR062 Liver (Clontech) Liver (Clontech) AR063 Lymphocytes chronic Lymphocytes chronic lymphocytic leukaemia lymphocytic leukaemia AR064 Lymphocytes diffuse Lymphocytes diffuse large B cell lymphoma large B cell lymphoma AR065 Lymphocytes follicular Lymphocytes follicular lymphoma lymphoma AR066 normal breast normal breast AR067 Normal Ovarian (4004901) Normal Ovarian (4004901) AR068 Normal Ovary 9508G045 Normal Ovary 9508G045 AR069 Normal Ovary 9701G208 Normal Ovary 9701G208 AR070 Normal Ovary 9806G005 Normal Ovary 9806G005 AR071 Ovarian Cancer Ovarian Cancer AR072 Ovarian Cancer (9702G001) Ovarian Cancer (9702G001) AR073 Ovarian Cancer (9707G029) Ovarian Cancer (9707G029) AR074 Ovarian Cancer (9804G011) Ovarian Cancer (9804G011) AR075 Ovarian Cancer (9806G019) Ovarian Cancer (9806G019) AR076 Ovarian Cancer (9807G017) Ovarian Cancer (9807G017) AR077 Ovarian Cancer (9809G001) Ovarian Cancer (9809G001) AR078 ovarian cancer 15799 ovarian cancer 15799 AR079 Ovarian Cancer 17717AID Ovarian Cancer 17717AID AR080 Ovarian Cancer 4004664B1 Ovarian Cancer 4004664B1 AR081 Ovarian Cancer 4005315A1 Ovarian Cancer 4005315A1 AR082 ovarian cancer 94127303 ovarian cancer 94127303 AR083 Ovarian Cancer 96069304 Ovarian Cancer 96069304 AR084 Ovarian Cancer 9707G029 Ovarian Cancer 9707G029 AR085 Ovarian Cancer 9807G045 Ovarian Cancer 9807G045 AR086 ovarian cancer 9809G001 ovarian cancer 9809G001 AR087 Ovarian Cancer 9905C032RC Ovarian Cancer 9905C032RC AR088 Ovarian cancer 9907 C00 3rd Ovarian cancer 9907 C00 3rd AR089 Prostate Prostate AR090 Prostate (clonetech) Prostate (clonetech) AR091 prostate cancer prostate cancer AR092 prostate cancer #15176 prostate cancer #15176 AR093 prostate cancer #15509 prostate cancer #15509 AR094 prostate cancer #15673 prostate cancer #15673 AR095 Small Intestine (Clontech) Small Intestine (Clontech) AR096 Spleen Spleen AR097 Thymus T cells activated Thymus T cells activated AR098 Thymus T cells resting Thymus T cells resting AR099 Tonsil Tonsil AR100 Tonsil geminal Tonsil geminal center centroblast center centroblast AR101 Tonsil germinal Tonsil germinal center B cell center B cell AR102 Tonsil lymph node Tonsil lymph node AR103 Tonsil memory B cell Tonsil memory B cell AR104 Whole Brain Whole Brain AR105 Xenograft ES-2 Xenograft ES-2 AR106 Xenograft SW626 Xenograft SW626 AR119 001: IL-2 001: IL-2 AR120 001: IL-2.1 001: IL-2.1 AR121 001: IL-2_b 001: IL-2_b AR124 002: Monocytes untreated 002: Monocytes untreated (1 hr) (1 hr) AR125 002: Monocytes untreated 002: Monocytes untreated (5 hrs) (5 hrs) AR126 002: Control.1C 002: Control.1C AR127 002: IL2.1C 002: IL2.1C AR130 003: Placebo-treated Rat 003: Placebo-treated Rat Lacrimal Gland Lacrimal Gland AR131 003: Placebo-treated Rat 003: Placebo-treated Rat Submandibular Gland Submandibular Gland AR135 004: Monocytes untreated (5 hrs) 004: Monocytes untreated (5 hrs) AR136 004: Monocytes untreated 1 hr 004: Monocytes untreated 1 hr AR139 005: Placebo (48 hrs) 005: Placebo (48 hrs) AR140 006: pC4 (24 hrs) 006: pC4 (24 hrs) AR141 006: pC4 (48 hrs) 006: pC4 (48 hrs) AR152 007: PHA(1 hr) 007: PHA(1 hr) AR153 007: PHA(6 HRS) 007: PHA(6 HRS) AR154 007: PMA(6 hrs) 007: PMA(6 hrs) AR155 008: 1449_’ 008: 1449_(#13) ’ AR161 01: A - max 24 01: A - max 24 AR162 01: A - max 26 01: A - max 26 AR163 01: A - max 30 01: A - max 30 AR164 01: B - max 24 01: B - max 24 AR165 01: B - max 26 01: B - max 26 AR166 01: B - max 30 01: B - max 30 AR167 1449 Sample 1449 Sample AR168 3T3P10 1.0 uM insulin 3T3P10 1.0 uM insulin AR169 3T3P10 10 nM Insulin 3T3P10 10 nM Insulin AR170 3T3P10 10 uM insulin 3T3P10 10 uM insulin AR171 3T3P10 No Insulin 3T3P10 No Insulin AR172 3T3P4 3T3P4 AR173 Adipose (41892) Adipose (41892) AR174 Adipose Diabetic (41611) Adipose Diabetic (41611) AR175 Adipose Diabetic (41661) Adipose Diabetic (41661) AR176 Adipose Diabetic (41689) Adipose Diabetic (41689) AR177 Adipose Diabetic (41706) Adipose Diabetic (41706) AR178 Adipose Diabetic (42352) Adipose Diabetic (42352) AR179 Adipose Diabetic (42366) Adipose Diabetic (42366) AR180 Adipose Diabetic (42452) Adipose Diabetic (42452) AR181 Adipose Diabetic (42491) Adipose Diabetic (42491) AR182 Adipose Normal (41843) Adipose Normal (41843) AR183 Adipose Normal (41893) Adipose Normal (41893) AR184 Adipose Normal (42452) Adipose Normal (42452) AR185 Adrenal Gland Adrenal Gland AR186 Adrenal Gland + Whole Brain Adrenal Gland + Whole Brain AR187 B7(1 hr) + (inverted) B7(1 hr) + (inverted) AR188 Breast (18275A2B) Breast (18275A2B) AR189 Breast (4004199) Breast (4004199) AR190 Breast (4004399) Breast (4004399) AR191 Breast (4004943B7) Breast (4004943B7) AR192 Breast (4005570B1) Breast (4005570B1) AR193 Breast Cancer (4004127A30) Breast Cancer (4004127A30) AR194 Breast Cancer (400443A21) Breast Cancer (400443A21) AR195 Breast Cancer (4004643A2) Breast Cancer (4004643A2) AR196 Breast Cancer (4004710A7) Breast Cancer (4004710A7) AR197 Breast Cancer (4004943A21) Breast Cancer (4004943A21) AR198 Breast Cancer (400553A2) Breast Cancer (400553A2) AR199 Breast Cancer (9805C046R) Breast Cancer (9805C046R) AR200 Breast Cancer (9806C012R) Breast Cancer (9806C012R) AR201 Breast Cancer (ODQ 45913) Breast Cancer (ODQ 45913) AR202 Breast Cancer (ODQ45913) Breast Cancer (ODQ45913) AR203 Breast Cancer (ODQ4591B) Breast Cancer (ODQ4591B) AR204 Colon Cancer (15663) Colon Cancer (15663) AR205 Colon Cancer (4005144A4) Colon Cancer (4005144A4) AR206 Colon Cancer (4005413A4) Colon Cancer (4005413A4) AR207 Colon Cancer (4005570B1) Colon Cancer (4005570B1) AR208 Control RNA #1 Control RNA #1 AR209 Control RNA #2 Control RNA #2 AR210 Cultured Preadipocyte (blue) Cultured Preadipocyte (blue) AR211 Cultured Preadipocyte (Red) Cultured Preadipocyte (Red) AR212 Donor II B-Cells 24 hrs Donor II B-Cells 24 hrs AR213 Donor II Resting B-Cells Donor II Resting B-Cells AR214 H114EP12 10 nM Insulin H114EP12 10 nM Insulin AR215 H114EP12 (10 nM insulin) H114EP12 (10 nM insulin) AR216 H114EP12 (2.6 ug/ul) H114EP12 (2.6 ug/ul) AR217 H114EP12 (3.6 ug/ul) H114EP12 (3.6 ug/ul) AR218 HUVEC #1 HUVEC #1 AR219 HUVEC #2 HUVEC #2 AR221 L6 undiff. L6 undiff. AR222 L6 Undifferentiated L6 Undifferentiated AR223 L6P8 + 10 nM Insulin L6P8 + 10 nM Insulin AR224 L6P8 + HS L6P8 + HS AR225 L6P8 10 nM Insulin L6P8 10 nM Insulin AR226 Liver (00-06-A007B) Liver (00-06-A007B) AR227 Liver (96-02-A075) Liver (96-02-A075) AR228 Liver (96-03-A144) Liver (96-03-A144) AR229 Liver (96-04-A138) Liver (96-04-A138) AR230 Liver (97-10-A074B) Liver (97-10-A074B) AR231 Liver (98-09-A242A) Liver (98-09-A242A) AR232 Liver Diabetic (1042) Liver Diabetic (1042) AR233 Liver Diabetic (41616) Liver Diabetic (41616) AR234 Liver Diabetic (41955) Liver Diabetic (41955) AR235 Liver Diabetic (42352R) Liver Diabetic (42352R) AR236 Liver Diabetic (42366) Liver Diabetic (42366) AR237 Liver Diabetic (42483) Liver Diabetic (42483) AR238 Liver Diabetic (42491) Liver Diabetic (42491) AR239 Liver Diabetic (99-09-A281A) Liver Diabetic (99-09-A281A) AR240 Lung Lung AR241 Lung (27270) Lung (27270) AR242 Lung (2727Q) Lung (2727Q) AR243 Lung Cancer (4005116A1) Lung Cancer (4005116A1) AR244 Lung Cancer (4005121A5) Lung Cancer (4005121A5) AR245 Lung Cancer (4005121A5)) Lung Cancer (4005121A5)) AR246 Lung Cancer (4005340A4) Lung Cancer (4005340A4) AR247 Mammary Gland Mammary Gland AR248 Monocyte (CT) Monocyte (CT) AR249 Monocyte (OCT) Monocyte (OCT) AR250 Monocytes (CT) Monocytes (CT) AR251 Monocytes (INFG 18 hr) Monocytes (INFG 18 hr) AR252 Monocytes (INFG 18 hr) Monocytes (INFG 18 hr) AR253 Monocytes (INFG 8-11) Monocytes (INFG 8-11) AR254 Monocytes (O CT) Monocytes (O CT) AR255 Muscle (91-01-A105) Muscle (91-01-A105) AR256 Muscle (92-04-A059) Muscle (92-04-A059) AR257 Muscle (97-11-A056d) Muscle (97-11-A056d) AR258 Muscle (99-06-A210A) Muscle (99-06-A210A) AR259 Muscle (99-07-A203B) Muscle (99-07-A203B) AR260 Muscle (99-7-A203B) Muscle (99-7-A203B) AR261 Muscle Diabetic (42352R) Muscle Diabetic (42352R) AR262 Muscle Diabetic (42366) Muscle Diabetic (42366) AR263 NK-19 Control NK-19 Control AR264 NK-19 IL Treated 72 hrs NK-19 IL Treated 72 hrs AR265 NK-19 UK Treated 72 hrs. NK-19 UK Treated 72 hrs. AR266 Omentum Normal (94-08-B009) Omentum Normal (94-08-B009) AR267 Omentum Normal (97-01-A039A) Omentum Normal (97-01-A039A) AR268 Omentum Normal (97-04-A114C) Omentum Normal (97-04-A114C) AR269 Omentum Normal (97-06-A117C) Omentum Normal (97-06-A117C) AR270 Omentum Normal (97-09-B004C) Omentum Normal (97-09-B004C) AR271 Ovarian Cancer (17717AID) Ovarian Cancer (17717AID) AR272 Ovarian Cancer (9905C023RC) Ovarian Cancer (9905C023RC) AR273 Ovarian Cancer (9905C032RC) Ovarian Cancer (9905C032RC) AR274 Ovary (9508G045) Ovary (9508G045) AR275 Ovary (9701G208) Ovary (9701G208) AR276 Ovary 9806G005 Ovary 9806G005 AR277 Pancreas Pancreas AR278 Placebo Placebo AR279 rIL2 Control rIL2 Control AR280 RSS288L RSS288L AR281 RSS288LC RSS288LC AR282 Salivary Gland Salivary Gland AR283 Skeletal Muscle Skeletal Muscle AR284 Skeletal Muscle (91-01-A105) Skeletal Muscle (91-01-A105) AR285 Skeletal Muscle (42180) Skeletal Muscle (42180) AR286 Skeletal Muscle (42386) Skeletal Muscle (42386) AR287 Skeletal Muscle (42461) Skeletal Muscle (42461) AR288 Skeletal Muscle (91-01-A105) Skeletal Muscle (91-01-A105) AR289 Skeletal Muscle (92-04-A059) Skeletal Muscle (92-04-A059) AR290 Skeletal Muscle (96-08-A171) Skeletal Muscle (96-08-A171) AR291 Skeletal Muscle (97-07-A190A) Skeletal Muscle (97-07-A190A) AR292 Skeletal Muscle Diabetic (42352) Skeletal Muscle Diabetic (42352) AR293 Skeletal Muscle Diabetic (42366) Skeletal Muscle Diabetic (42366) AR294 Skeletal Muscle Diabetic (42395) Skeletal Muscle Diabetic (42395) AR295 Skeletal Muscle Diabetic (42483) Skeletal Muscle Diabetic (42483) AR296 Skeletal Muscle Diabetic (42491) Skeletal Muscle Diabetic (42491) AR297 Skeletal Muscle Diabetic 42352 Skeletal Muscle Diabetic 42352 AR298 Skeletal Musle (42461) Skeletal Musle (42461) AR299 Small Intestine Small Intestine AR300 Stomach Stomach AR301 T-Cell + HDPBQ71.fc 1449 T-Cell + HDPBQ71.fc 1449 16 hrs 16 hrs AR302 T-Cell + HDPBQ71.fc 1449 T-Cell + HDPBQ71.fc 1449 6 hrs 6 hrs AR303 T-Cell + IL2 16 hrs T-Cell + IL2 16 hrs AR304 T-Cell + IL2 6 hrs T-Cell + IL2 6 hrs AR306 T-Cell Untreated 16 hrs T-Cell Untreated 16 hrs AR307 T-Cell Untreated 6 hrs T-Cell Untreated 6 hrs AR308 T-Cells 24 hours T-Cells 24 hours AR309 T-Cells 24 hrs T-Cells 24 hrs AR310 T-Cells 24 hrs. T-Cells 24 hrs. AR311 T-Cells 24 hrs T-Cells 24 hrs AR312 T-Cells 4 days T-Cells 4 days AR313 Thymus Thymus AR314 TRE TRE AR315 TREC TREC AR316 Virtual Mixture Virtual Mixture AR317 B lymphocyte, B lymphocyte, AR318 (non-T; non-B) (non-T; non-B) AR326 001 - 293 RNA (Vector Control) 001 - 293 RNA (Vector Control) AR327 001: Control 001: Control AR328 001: Control.1 001: Control. 1 AR355 Acute Lymphocyte Leukemia Acute Lymphocyte Leukemia AR356 AML Patient #11 AML Patient #11 AR357 AML Patient #2 AML Patient #2 AR358 AML Patient #2 SGAH AML Patient #2 SGAH AR359 AML Patient#2 AML Patient#2 AR360 Aorta Aorta AR361 B Cell B Cell AR362 B lymphoblast B lymphoblast AR363 B lymphocyte B lymphocyte AR364 B lymphocytes B lymphocytes AR365 B-cell B-cell AR366 B-Cells B-Cells AR367 B-Lymphoblast B-Lymphoblast AR368 B-Lymphocytes B-Lymphocytes AR369 Bladder Bladder AR370 Bone Marrow Bone Marrow AR371 Bronchial Epithelial Cell Bronchial Epithelial Cell AR372 Bronchial Epithelial Cells Bronchial Epithelial Cells AR373 Caco-2A Caco-2A AR374 Caco-2B Caco-2B AR375 Caco-2C Caco-2C AR376 Cardiac #1 Cardiac #1 AR377 Cardiac #2 Cardiac #2 AR378 Chest Muscle Chest Muscle AR381 Dendritic Cell Dendritic Cell AR382 Dendritic cells Dendritic cells AR383 E. coli E. coli AR384 Epithelial Cells Epithelial Cells AR385 Esophagus Esophagus AR386 FPPS FPPS AR387 FPPSC FPPSC AR388 HepG2 Cell Line HepG2 Cell Line AR389 HepG2 Cell line Buffer 1 hr. HepG2 Cell line Buffer 1 hr. AR390 HepG2 Cell line Buffer 06 hr HepG2 Cell line Buffer 06 hr AR391 HepG2 Cell line Buffer 24 hr. HepG2 Cell line Buffer 24 hr. AR392 HepG2 Cell line Insulin 01 hr. HepG2 Cell line Insulin 01 hr. AR393 HepG2 Cell line Insulin 06 hr. HepG2 Cell line Insulin 06 hr. AR394 HepG2 Cell line Insulin 24 hr. HepG2 Cell line Insulin 24 hr. AR398 HMC-1 HMC-1 AR399 HMCS HMCS AR400 HMSC HMSC AR401 HUVEC #3 HUVEC #3 AR402 HUVEC #4 HUVEC #4 AR404 KIDNEY NORMAL KIDNEY NORMAL AR405 KIDNEY TUMOR KIDNEY TUMOR AR406 KIDNEY TUMOR AR407 Lymph Node Lymph Node AR408 Macrophage Macrophage AR409 Megakarioblast Megakarioblast AR410 Monocyte Monocyte AR411 Monocytes Monocytes AR412 Myocardium Myocardium AR413 Myocardium #3 Myocardium #3 AR414 Myocardium #4 Myocardium #4 AR415 Myocardium #5 Myocardium #5 AR416 NK NK AR417 NK cell NK cell AR418 NK cells NK cells AR419 NKYa NKYa AR420 NKYa019 NKYa019 AR421 Ovary Ovary AR422 Patient #11 Patient #11 AR423 Peripheral blood Peripheral blood AR424 Primary Adipocytes Primary Adipocytes AR425 Promyeloblast Promyeloblast AR427 RSSWT RSSWT AR428 RSSWTC RSSWTC AR429 SW 480(G1) SW 480(G1) AR430 SW 480(G2) SW 480(G2) AR431 SW 480(G3) SW 480(G3) AR432 SW 480(G4) SW 480(G4) AR433 SW 480(G5) SW 480(G5) AR434 T Lymphoblast T Lymphoblast AR435 T Lymphocyte T Lymphocyte AR436 T-Cell T-Cell AR438 T-Cell, T-Cell, AR439 T-Cells T-Cells AR440 T-lymphoblast T-lymphoblast AR441 Th 1 Th 1 AR442 Th 2 Th 2 AR443 Th1 Th1 AR444 Th2 Th2 H0002 Human Adult Heart Human Adult Heart Heart Uni-ZAP XR H0003 Human Adult Liver Human Adult Liver Liver Uni-ZAP XR H0004 Human Adult Spleen Human Adult Spleen Spleen Uni-ZAP XR H0007 Human Cerebellum Human Cerebellum Brain Uni-ZAP XR H0008 Whole 6 Week Old Embryo Uni-ZAP XR H0009 Human Fetal Brain Uni-ZAP XR H0010 Human Fetal Hepatic Human Fetal Liver Liver Uni-ZAP XR H0011 Human Fetal Kidney Human Fetal Kidney Kidney Uni-ZAP XR H0012 Human Fetal Kidney Human Fetal Kidney Kidney Uni-ZAP XR H0013 Human 8 Week Whole Embryo Human 8 Week Old Embryo Embryo Uni-ZAP XR H0014 Human Gall Bladder Human Gall Bladder Gall Bladder Uni-ZAP XR H0015 Human Gall Bladder, fraction II Human Gall Bladder Gall Bladder Uni-ZAP XR H0018 Human Greater Omentum, fII remake Human Greater Omentum peritoneum Uni-ZAP XR H0019 Human Fetal Heart Human Fetal Heart Heart pBluescript H0020 Human Hippocampus Human Hippocampus Brain Uni-ZAP XR H0021 Human Infant Adrenal Gland Human Infant Adrenal Gland Adrenal gland Uni-ZAP XR H0022 Jurkat Cells Jurkat T-Cell Line Lambda ZAP II H0023 Human Fetal Lung Uni-ZAP XR H0024 Human Fetal Lung III Human Fetal Lung Lung Uni-ZAP XR H0025 Human Adult Lymph Node Human Adult Lymph Node Lymph Node Lambda ZAP II H0026 Namalwa Cells Namalwa B-Cell Line, Lambda ZAP II EBV immortalized H0028 Human Old Ovary Human Old Ovary Ovary pBluescript H0029 Human Pancreas Human Pancreas Pancreas Uni-ZAP XR H0030 Human Placenta Uni-ZAP XR H0031 Human Placenta Human Placenta Placenta Uni-ZAP XR H0032 Human Prostate Human Prostate Prostate Uni-ZAP XR H0033 Human Pituitary Human Pituitary Uni-ZAP XR H0035 Human Salivary Gland Human Salivary Gland Salivary gland Uni-ZAP XR H0036 Human Adult Small Intestine Human Adult Small Intestine Small Int. Uni-ZAP XR H0037 Human Adult Small Intestine Human Adult Small Intestine Small Int. pBluescript H0038 Human Testes Human Testes Testis Uni-ZAP XR H0039 Human Pancreas Tumor Human Pancreas Tumor Pancreas disease Uni-ZAP XR H0040 Human Testes Tumor Human Testes Tumor Testis disease Uni-ZAP XR H0041 Human Fetal Bone Human Fetal Bone Bone Uni-ZAP XR H0042 Human Adult Pulmonary Human Adult Pulmonary Lung Uni-ZAP XR H0044 Human Cornea Human Cornea eye Uni-ZAP XR H0045 Human Esophagus, Cancer Human Esophagus, cancer Esophagus disease Uni-ZAP XR H0046 Human Endometrial Tumor Human Endometrial Tumor Uterus disease Uni-ZAP XR H0047 Human Fetal Liver Human Fetal Liver Liver Uni-ZAP XR H0048 Human Pineal Gland Human Pineal Gland Uni-ZAP XR H0049 Human Fetal Kidney Human Fetal Kidney Kidney Uni-ZAP XR H0050 Human Fetal Heart Human Fetal Heart Heart Uni-ZAP XR H0051 Human Hippocampus Human Hippocampus Brain Uni-ZAP XR H0052 Human Cerebellum Human Cerebellum Brain Uni-ZAP XR H0053 Human Adult Kidney Human Adult Kidney Kidney Uni-ZAP XR H0056 Human Umbilical Human Umbilical Vein Umbilical Uni-ZAP XR Vein, Endo. remake Endothelial Cells vein H0057 Human Fetal Spleen Uni-ZAP XR H0058 Human Thymus Tumor Human Thymus Tumor Thymus disease Lambda ZAP II H0059 Human Uterine Cancer Human Uterine Cancer Uterus disease Lambda ZAP II H0060 Human Macrophage Human Macrophage Blood Cell Line pBluescript H0061 Human Macrophage Human Macrophage Blood Cell Line pBluescript H0062 Human Thymus Human Thymus Thymus Uni-ZAP XR H0063 Human Thymus Human Thymus Thymus Uni-ZAP XR H0064 Human Right Hemisphere of Brain Human Brain, right Brain Uni-ZAP XR hemisphere H0067 Human left hemisphere, adult Human Left Hemisphere, Adult Brain Lambda ZAP II H0068 Human Skin Tumor Human Skin Tumor Skin disease Uni-ZAP XR H0069 Human Activated T-Cells Activated T-Cells Blood Cell Line Uni-ZAP XR H0070 Human Pancreas Human Pancreas Pancreas Uni-ZAP XR H0071 Human Infant Adrenal Gland Human Infant Adrenal Gland Adrenal gland Uni-ZAP XR H0073 Human Leiomyeloid Carcinoma Human Leiomyeloid Carcinoma Muscle disease Uni-ZAP XR H0074 Human Platelets Human Platelets Blood Cell Line Uni-ZAP XR H0075 Human Activated T-Cells (II) Activated T-Cells Blood Cell Line Uni-ZAP XR H0077 Human Thymus Tumor Human Thymus Tumor Thymus disease Lambda-ZAP II H0078 Human Lung Cancer Human Lung Cancer Lung disease Lambda ZAP II H0079 Human Whole 7 Week Old Embryo (II) Human Whole 7 Week Old Embryo Embryo Uni-ZAP XR H0080 Human Whole 6 Week Old Embryo (II) Human Whole Six Week Old Embryo Lambda ZAP II Embryo H0081 Human Fetal Epithelium Human Fetal Skin Skin Uni-ZAP XR (Skin) H0082 Human Fetal Muscle Human Fetal Muscle Sk Muscle Uni-ZAP XR H0083 HUMAN JURKAT Jurkat Cells Uni-ZAP XR MEMBRANE BOUND POLYSOMES H0085 Human Colon Human Colon Lambda ZAP II H0086 Human epithelioid sarcoma Epithelioid Sk Muscle disease Uni-ZAP XR Sarcoma, muscle H0087 Human Thymus Human Thymus pBluescript H0090 Human T-Cell Lymphoma T-Cell Lymphoma T-Cell disease Uni-ZAP XR H0092 Human Pancreas Tumor Human Pancreas Tumor Pancreas disease Uni-ZAP XR H0093 Human Greater Omentum Tumor Human Greater Omentum peritoneum disease Uni-ZAP XR H0095 Human Greater Omentum, RNA Remake Human Greater Omentum peritoneum Uni-ZAP XR H0097 Human Adult Heart, subtracted Human Adult Heart Heart pBluescript H0098 Human Adult Liver, subtracted Human Adult Liver Liver Uni-ZAP XR H0099 Human Lung Cancer, subtracted Human Lung Cancer Lung pBluescript H0100 Human Whole Six Week Human Whole Six Embryo Uni-ZAP XR Old Embryo Week Old Embryo H0101 Human 7 Weeks Old Embryo, subtracted Human Whole 7 Week Old Embryo Lambda ZAP II Embryo H0102 Human Whole 6 Week Old Human Whole Six Week Old Embryo pBluescript Embryo (II), subt Embryo H0103 Human Fetal Brain, subtracted Human Fetal Brain Brain Uni-ZAP XR H0105 Human Fetal Heart, subtracted Human Fetal Heart Heart pBluescript H0107 Human Infant Adrenal Gland, subtracted Human Infant Adrenal Gland Adrenal gland pBluescript H0108 Human Adult Lymph Node, subtracted Human Adult Lymph Node Lymph Node Uni-ZAP XR H0109 Human Macrophage, subtracted Macrophage Blood Cell Line pBluescript H0110 Human Old Ovary, subtracted Human Old Ovary Ovary pBluescript H0111 Human Placenta, subtracted Human Placenta Placenta pBluescript H0113 Human skin Tumor, subtracted Human Skin Tumor Skin Uni-ZAP XR H0116 Human Thymus Tumor, subtracted Human Thymus Tumor Thymus pBluescript H0117 Human Uterine Cancer, subtracted Human Uterine Cancer Uterus pBluescript H0119 Human Pediatric Kidney Human Pediatric Kidney Kidney Uni-ZAP XR H0120 Human Adult Spleen, subtracted Human Adult Spleen Spleen Uni-ZAP XR H0121 Human Cornea, subtracted Human Cornea eye Uni-ZAP XR H0122 Human Adult Skeletal Muscle Human Skeletal Muscle Sk Muscle Uni-ZAP XR H0123 Human Fetal Dura Mater Human Fetal Dura Mater Brain Uni-ZAP XR H0124 Human Rhabdomyosarcoma Human Rhabdomyosarcoma Sk Muscle disease Uni-ZAP XR H0125 Cem cells cyclohexamide Cyclohexamide Treated Cem, Blood Cell Line Uni-ZAP XR treated Jurkat, Raji, and Supt H0128 Jurkat cells, thiouridine activated Jurkat Cells Uni-ZAP XR H0129 Jurkat cells, thiouridine activated, fract II Jurkat Cells Uni-ZAP XR H0130 LNCAP untreated LNCAP Cell Line Prostate Cell Line Uni-ZAP XR H0131 LNCAP + o.3 nM R1881 LNCAP Cell Line Prostate Cell Line Uni-ZAP XR H0132 LNCAP + 30 nM R1881 LNCAP Cell Line Prostate Cell Line Uni-ZAP XR H0134 Raji Cells, cyclohexamide Cyclohexamide Treated Cem, Blood Cell Line Uni-ZAP XR treated Jurkat, Raji, and Supt H0135 Human Synovial Sarcoma Human Synovial Sarcoma Synovium Uni-ZAP XR H0136 Supt Cells, cyclohexamide treated Cyclohexamide Treated Cem, Blood Cell Line Uni-ZAP XR Jurkat, Raji, and Supt H0140 Activated T-Cells, 8 hrs. Activated T-Cells Blood Cell Line Uni-ZAP XR H0141 Activated T-Cells, 12 hrs. Activated T-Cells Blood Cell Line Uni-ZAP XR H0142 MCF7 Cell Line MCF7 Cell line Breast Cell Line Uni-ZAP XR H0144 Nine Week Old Early 9 Wk Old Early Embryo Uni-ZAP XR Stage Human Stage Human H0147 Human Adult Liver Human Adult Liver Liver Uni-ZAP XR H0149 7 Week Old Early Stage Human, Human Whole 7 Week Old Embryo Uni-ZAP XR subtracted Embryo H0150 Human Epididymus Epididymis Testis Uni-ZAP XR H0151 Early Stage Human Liver Human Fetal Liver Liver Uni-ZAP XR H0152 Early Stage Human Liver, fract (II) Human Fetal Liver Liver Uni-ZAP XR H0154 Human Fibrosarcoma Human Skin Fibrosarcoma Skin disease Uni-ZAP XR H0155 Human Thymus, subtracted Human Thymus Tumor Thymus pBluescript H0156 Human Adrenal Gland Tumor Human Adrenal Gland Tumor Adrenal Gland disease Uni-ZAP XR H0157 Activated T-Cells, 0 hrs, ligation 2 Activated T-Cells Blood Cell Line Uni-ZAP XR H0158 Activated T-Cells, 4 hrs., ligation 2 Activated T-Cells Blood Cell Line Uni-ZAP XR H0159 Activated T-Cells, 8 hrs., ligation 2 Activated T-Cells Blood Cell Line Uni-ZAP XR H0163 Human Synovium Human Synovium Synovium Uni-ZAP XR H0164 Human Trachea Tumor Human Trachea Tumor Trachea disease Uni-ZAP XR H0165 Human Prostate Cancer, Human Prostate Prostate disease Uni-ZAP XR Stage B2 Cancer, stage B2 H0166 Human Prostate Cancer, Human Prostate Prostate disease Uni-ZAP XR Stage B2 Cancer, stage B2 H0167 Activated T-Cells, Activated T-Cells Blood Cell Line Uni-ZAP XR 24 hrs. H0168 Human Prostate Cancer, Stage C Human Prostate Cancer, Prostate disease Uni-ZAP XR stage C H0169 Human Prostate Cancer, Stage C Human Prostate Cancer, Prostate disease Uni-ZAP XR fraction stage C H0170 12 Week Old Early Stage Human Twelve Week Old Embryo Uni-ZAP XR Early Stage Human H0171 12 Week Old Early Stage Twelve Week Old Embryo Uni-ZAP XR Human, II Early Stage Human H0172 Human Fetal Brain, random primed Human Fetal Brain Brain Lambda ZAP II H0173 Human Cardiomyopathy, RNA remake Human Cardiomyopathy Heart disease Uni-ZAP XR H0175 H. Adult Spleen, ziplox pSport1 H0176 CAMA1Ee Cell Line CAMA1Ee Cell Line Breast Cell Line Uni-ZAP XR H0177 CAMA1Ee Cell Line CAMA1Ee Cell Line Breast Cell Line Uni-ZAP XR H0178 Human Fetal Brain Human Fetal Brain Brain Uni-ZAP XR H0179 Human Neutrophil Human Neutrophil Blood Cell Line Uni-ZAP XR H0181 Human Primary Breast Cancer Human Primary Breast disease Uni-ZAP XR Breast Cancer H0182 Human Primary Breast Cancer Human Primary Breast Breast disease Uni-ZAP XR Cancer H0183 Human Colon Cancer Human Colon Cancer Colon disease Uni-ZAP XR H0184 Human Colon Cancer, metasticized to live Human Colon Cancer, Liver disease Lambda ZAP II metasticized to liver H0185 Activated T-Cell labeled with 4-thioluri T-Cells Blood Cell Line Lambda ZAP II H0186 Activated T-Cell T-Cells Blood Cell Line Lambda ZAP II H0187 Resting T-Cell T Cells Blood Cell Line Lambda ZAP II H0188 Human Normal Breast Human Normal Breast Breast Uni-ZAP XR H0189 Human Resting Macrophage Human Macrophage/Monocytes Blood Cell Line Uni-ZAP XR H0190 Human Activated Macrophage (LPS) Human Macrophage/Monocytes Blood Cell Line Uni-ZAP XR H0191 Human Activated Macrophage (LPS), thiour Human Macrophage/Monocytes Blood Cell Line Uni-ZAP XR H0192 Cem Cells, cyclohexamide treated, subtra Cyclohexamide Treated Blood Cell Line Uni-ZAP XR Cem, Jurkat, Raji, and Supt H0193 Cem Cells, cyclohexamide treated, differ Cyclohexamide Treated Blood Cell Line Uni-ZAP XR Cem, Jurkat, Raji, and Supt H0194 Human Cerebellum, subtracted Human Cerebellum Brain pBluescript H0196 Human Cardiomyopathy, subtracted Human Cardiomyopathy Heart Uni-ZAP XR H0197 Human Fetal Liver, subtracted Human Fetal Liver Liver Uni-ZAP XR H0198 Human Fetal Liver, subtracted, pos. clon Human Fetal Liver Liver Uni-ZAP XR H0199 Human Fetal Liver, subtracted, neg clone Human Fetal Liver Liver Uni-ZAP XR H0200 Human Greater Omentum, fract II remake, Human Greater Omentum peritoneum Uni-ZAP XR H0201 Human Hippocampus, subtracted Human Hippocampus Brain pBluescript H0202 Jurkat Cells, cyclohexamide treated, Cyclohexamide Treated Blood Cell Line Uni-ZAP XR subtraction Cem, Jurkat, Raji, and Supt H0203 Jurkat Cells, cyclohexamide treated, dif Cyclohexamide Treated Blood Cell Line Uni-ZAP XR Cem, Jurkat, Raji, and Supt H0204 Human Colon Cancer, subtracted Human Colon Cancer Colon pBluescript H0205 Human Colon Cancer, differential Human Colon Cancer Colon pBluescript H0207 LNCAP, differential expression LNCAP Cell Line Prostate Cell Line pBluescript H0208 Early Stage Human Lung, Human Fetal Lung Lung pBluescript subtracted H0209 Human Cerebellum, differentially expressed Human Cerebellum Brain Uni-ZAP XR H0211 Human Prostate, differential expression Human Prostate Prostate pBluescript H0212 Human Prostate, subtracted Human Prostate Prostate pBluescript H0213 Human Pituitary, subtracted Human Pituitary Uni-ZAP XR H0214 Raji cells, cyclohexamide treated, subtracted Cyclohexamide Treated Blood Cell Line pBluescript Cem, Jurkat, Raji, and Supt H0215 Raji cells, cyclohexamide treated, Cyclohexamide Treated Blood Cell Line pBluescript differentially expressed Cem, Jurkat, Raji, and Supt H0216 Supt cells, cyclohexamide treated, subtracted Cyclohexamide Treated Blood Cell Line pBluescript Cem, Jurkat, Raji, and Supt H0217 Supt cells, cyclohexamide treated, Cyclohexamide Treated Cem, Blood Cell Line pBluescript differentially expressed Jurkat, Raji, and Supt H0218 Activated T-Cells, 0 hrs, subtracted Activated T-Cells Blood Cell Line Uni-ZAP XR H0219 Activated T-Cells, 0 hrs, differentially Activated T-Cells Blood Cell Line Uni-ZAP XR expressed H0220 Activated T-Cells, 4 hrs, subtracted Activated T-Cells Blood Cell Line Uni-ZAP XR H0222 Activated T-Cells, 8 hrs, subtracted Activated T-Cells Blood Cell Line Uni-ZAP XR H0224 Activated T-Cells, 12 hrs, subtracted Activated T-Cells Blood Cell Line Uni-ZAP XR H0225 Activated T-Cells, 12 hrs, differentially Activated T-Cells Blood Cell Line Uni-ZAP XR expressed H0228 C7MCF7 cell line, estrogen treated C7MCF7 Cell Line, estrogen Breast Cell Line Uni-ZAP XR treated H0229 Early Stage Human Brain, random primed Early Stage Human Brain Brain Lambda ZAP II H0230 Human Cardiomyopathy, diff exp Human Cardiomyopathy Heart disease Uni-ZAP XR H0231 Human Colon, subtraction Human Colon pBluescript H0232 Human Colon, differential expression Human Colon pBluescript H0233 Human Fetal Heart, Differential (Adult- Human Fetal Heart Heart pBluescript Specific) H0234 human colon cancer, metastatic Human Colon Cancer, Liver pBluescript to liver, differentially expressed metasticized to liver H0235 Human colon cancer, metaticized to liver, Human Colon Cancer, Liver pBluescript subtraction metasticized to liver H0239 Human Kidney Tumor Human Kidney Tumor Kidney disease Uni-ZAP XR H0241 C7MCF7 cell line, estrogen treated, C7MCF7 Cell Line, estrogen Breast Cell Line Uni-ZAP XR subtraction treated H0242 Human Fetal Heart, Differential (Fetal- Human Fetal Heart Heart pBluescript Specific) H0244 Human 8 Week Whole Embryo, Human 8 Week Whole Embryo Embryo Uni-ZAP XR subtracted H0246 Human Fetal Liver- Enzyme Human Fetal Liver Liver Uni-ZAP XR subtraction H0247 Human Membrane Bound Polysomes- Human Membrane Bound Blood Cell Line Uni-ZAP XR Enzyme Subtraction Polysomes H0249 HE7, subtracted by hybridization with E7 Human Whole 7 Week Old Embryo Uni-ZAP XR cDNA Embryo H0250 Human Activated Human Monocytes Uni-ZAP XR Monocytes H0251 Human Chondrosarcoma Human Chondrosarcoma Cartilage disease Uni-ZAP XR H0252 Human Osteosarcoma Human Osteosarcoma Bone disease Uni-ZAP XR H0253 Human adult testis, large inserts Human Adult Testis Testis Uni-ZAP XR H0254 Breast Lymph node cDNA library Breast Lymph Node Lymph Node Uni-ZAP XR H0255 breast lymph node CDNA library Breast Lymph Node Lymph Node Lambda ZAP II H0256 HL-60, unstimulated Human HL-60 Cells, Blood Cell Line Uni-ZAP XR unstimulated H0257 HL-60, PMA 4 H HL-60 Cells, PMA Blood Cell Line Uni-ZAP XR stimulated 4 H H0261 H. cerebellum, Enzyme subtracted Human Cerebellum Brain Uni-ZAP XR H0263 human colon cancer Human Colon Cancer Colon disease Lambda ZAP II H0264 human tonsils Human Tonsil Tonsil Uni-ZAP XR H0265 Activated T-Cell T-Cells Blood Cell Line Uni-ZAP XR (12 hs)/Thiouridine labelledEco H0266 Human Microvascular HMEC Vein Cell Line Lambda ZAP II Endothelial Cells, fract. A H0267 Human Microvascular Endothelial Cells, HMEC Vein Cell Line Lambda ZAP II fract. B H0268 Human Umbilical Vein HUVE Cells Umbilical Cell Line Lambda ZAP II Endothelial Cells, fract. A vein H0269 Human Umbilical Vein Endothelial Cells, HUVE Cells Umbilical Cell Line Lambda ZAP II fract. B vein H0270 HPAS (human pancreas, subtracted) Human Pancreas Pancreas Uni-ZAP XR H0271 Human Neutrophil, Activated Human Neutrophil - Activated Blood Cell Line Uni-ZAP XR H0272 HUMAN TONSILS, FRACTION 2 Human Tonsil Tonsil Uni-ZAP XR H0274 Human Adult Spleen, fractionII Human Adult Spleen Spleen Uni-ZAP XR H0275 Human Infant Adrenal Gland, Human Infant Adrenal Gland Adrenal gland pBluescript Subtracted H0279 K562 cells K562 Cell line cell line Cell Line ZAP Express H0280 K562 + PMA (36 hrs) K562 Cell line cell line Cell Line ZAP Express H0281 Lymph node, abnorm. cell line (ATCC Lymph Node, abnormal cell Lymph Node Cell Line ZAP Express #7225) line H0282 HBGB''s differential consolidation Human Primary Breast Cancer Breast Uni-ZAP XR H0284 Human OB MG63 control Human Osteoblastoma Bone Cell Line Uni-ZAP XR fraction I MG63 cell line H0286 Human OB MG63 treated Human Osteoblastoma Bone Cell Line Uni-ZAP XR (10 nM E2) fraction I MG63 cell line H0288 Human OB HOS control fraction I Human Osteoblastoma Bone Cell Line Uni-ZAP XR HOS cell line H0290 Human OB HOS treated (1 nM E2) fraction I Human Osteoblastoma HOS Bone Cell Line Uni-ZAP XR cell line H0292 Human OB HOS treated (10 nM E2) fraction I Human Osteoblastoma HOS Bone Cell Line Uni-ZAP XR cell line H0293 WI 38 cells Uni-ZAP XR H0294 Amniotic Cells - TNF induced Amniotic Cells - Placenta Cell Line Uni-ZAP XR TNF induced H0295 Amniotic Cells- Primary Culture Amniotic Cells - Placenta Cell Line Uni-ZAP XR Primary Culture H0298 HCBB''s differential consolidation CAMA1Ee Cell Line Breast Cell Line Uni-ZAP XR H0299 HCBA''s differential consolidation CAMA1Ee Cell Line Breast Cell Line Uni-ZAP XR H0300 CD34 positive cells (Cord Blood) CD34 Positive Cells Cord Blood ZAP Express H0305 CD34 positive cells (Cord Blood) CD34 Positive Cells Cord Blood ZAP Express H0306 CD34 depleted Buffy Coat CD34 Depleted Cord Blood ZAP Express (Cord Blood) Buffy Coat (Cord Blood) H0309 Human Chronic Synovitis Synovium, Chronic Synovium disease Uni-ZAP XR Synovitis/Osteoarthritis H0310 human caudate nucleus Brain Brain Uni-ZAP XR H0313 human pleural cancer pleural cancer disease pBluescript H0316 HUMAN STOMACH Human Stomach Stomach Uni-ZAP XR H0318 HUMAN B CELL LYMPHOMA Human B Cell Lymphoma Lymph Node disease Uni-ZAP XR H0320 Human frontal cortex Human Frontal Cortex Brain Uni-ZAP XR H0321 HUMAN SCHWANOMA Schwanoma Nerve disease Uni-ZAP XR H0327 human corpus colosum Human Corpus Callosum Brain Uni-ZAP XR H0328 human ovarian cancer Ovarian Cancer Ovary disease Uni-ZAP XR H0329 Dermatofibrosarcoma Dermatofibrosarcoma Skin disease Uni-ZAP XR Protuberance Protuberans H0330 HCBB''s Subtractive (−mito genes) CAMA1Ee Cell Line Breast Cell Line Uni-ZAP XR H0331 Hepatocellular Tumor Hepatocellular Tumor Liver disease Lambda ZAP II H0333 Hemangiopericytoma Hemangiopericytoma Blood vessel disease Lambda ZAP II H0334 Kidney cancer Kidney Cancer Kidney disease Uni-ZAP XR H0339 Duodenum Duodenum Uni-ZAP XR H0340 Corpus Callosum Corpus Collosum-93052 Uni-ZAP XR H0341 Bone Marrow Cell Line Bone Marrow Cell Bone Marrow Cell Line Uni-ZAP XR (RS4; 11) Line RS4; 11 H0342 Lingual Gyrus Lingual Gyrus Brain Uni-Zap XR H0343 stomach cancer (human) Stomach Cancer - 5383A disease Uni-ZAP XR (human) H0344 Adipose tissue (human) Adipose - 6825A (human) Uni-ZAP XR H0345 SKIN Skin - 4000868H Skin Uni-ZAP XR H0346 Brain-medulloblastoma Brain (Medulloblastoma)- Brain disease Uni-ZAP XR 9405C006R H0349 human adult liver cDNA library Human Adult Liver Liver pCMVSport 1 H0350 Human Fetal Liver, mixed 10 & 14 week Human Fetal Liver, mixed Liver Uni-ZAP XR 10&14 Week H0351 Glioblastoma Glioblastoma Brain disease Uni-ZAP XR H0352 wilm''s tumor Wilm''s Tumor disease Uni-ZAP XR H0354 Human Leukocytes Human Leukocytes Blood Cell Line pCMVSport 1 H0355 Human Liver Human Liver, normal Adult pCMVSport 1 H0356 Human Kidney Human Kidney Kidney pCMVSport 1 H0357 H. Normalized Fetal Liver, II Human Fetal Liver Liver Uni-ZAP XR H0359 KMH2 cell line KMH2 ZAP Express H0360 Hemangiopericytoma Hemangiopericytoma disease H0361 Human rejected kidney Human Rejected Kidney disease pBluescript H0362 HeLa cell line HELA CELL LINE pSport1 H0363 Human Brain Medulla, subtracted Human Brain Medulla pBluescript H0364 Human Osteoclastoma, excised Human Osteoclastoma disease pBluescript H0365 Osteoclastoma-normalized B Human Osteoclastoma disease Uni-ZAP XR H0366 L428 cell line L428 ZAP Express H0369 H. Atrophic Endometrium Atrophic Endometrium and Uni-ZAP XR myometrium H0370 H. Lymph node breast Cancer Lymph node with disease Uni-ZAP XR Met. Breast Cancer H0371 Eosinophils-Hypereosinophilia patient Eosinophils- disease Uni-ZAP XR Hypereosinophilia patient H0372 Human Testes Human Testes Testis pCMVSport 1 H0373 Human Heart Human Adult Heart Heart pCMVSport 1 H0374 Human Brain Human Brain pCMVSport 1 H0375 Human Lung Human Lung pCMVSport 1 H0376 Human Spleen Human Adult Spleen Spleen pCMVSport 1 H0379 Human Tongue, frac 1 Human Tongue pSport1 H0380 Human Tongue, frac 2 Human Tongue pSport1 H0381 Bone Cancer Bone Cancer disease Uni-ZAP XR H0383 Human Prostate BPH, re-excision Human Prostate BPH Uni-ZAP XR H0384 Brain, Kozak Human Brain pCMVSport 1 H0385 H. Leukocytes, Kozak Human Leukocytes Blood Cell Line pCMVSport 1 H0386 Leukocyte and Lung; 4 screens Human Leukocytes Blood Cell Line pCMVSport 1 H0388 Human Rejected Kidney, 704 re-excision Human Rejected Kidney disease pBluescript H0389 H. Brain, X-Chromosome hybridization Human Brain pCMVSport 1 H0390 Human Amygdala Depression, re-excision Human Amygdala Depression disease pBluescript H0391 H. Meningima, M6 Human Meningima brain pSport1 H0392 H. Meningima, M1 Human Meningima brain pSport1 H0393 Fetal Liver, subtraction II Human Fetal Liver Liver pBluescript H0394 A-14 cell line Redd-Sternberg cell ZAP Express H0395 A1-CELL LINE Redd-Sternberg cell ZAP Express H0396 L1 Cell line Redd-Sternberg cell ZAP Express H0398 Human Newborn Bladder Human Newborn Bladder pBluescript H0399 Human Kidney Cortex, re-rescue Human Kidney Cortex Lambda ZAP II H0400 Human Striatum Depression, re-rescue Human Brain, Striatum Brain Lambda ZAP II Depression H0401 Human Pituitary, subtracted V Human Pituitary pBluescript H0402 CD34 depleted Buffy Coat CD34 Depleted Cord Blood ZAP Express (Cord Blood), re-excision Buffy Coat (Cord Blood) H0403 H. Umbilical Vein Endothelial Cells, IL4 HUVE Cells Umbilical Cell Line Uni-ZAP XR induced vein H0404 H. Umbilical Vein endothelial cells, HUVE Cells Umbilical Cell Line Uni-ZAP XR uninduced vein H0405 Human Pituitary, subtracted VI Human Pituitary pBluescript H0406 H Amygdala Depression, subtracted Human Amygdala Uni-ZAP XR Depression H0408 Human kidney Cortex, subtracted Human Kidney Cortex pBluescript H0409 H. Striatum Depression, subtracted Human Brain, Striatum Brain pBluescript Depression H0410 H. Male bladder, adult H Male Bladder, Adult Bladder pSport1 H0411 H Female Bladder, Adult Human Female Adult Bladder Bladder pSport1 H0412 Human umbilical vein endothelial HUVE Cells Umbilical Cell Line pSport1 cells, IL-4 induced vein H0413 Human Umbilical Vein Endothelial HUVE Cells Umbilical Cell Line pSport1 Cells, uninduced vein H0414 Ovarian Tumor I, OV5232 Ovarian Tumor, OV5232 Ovary disease pSport1 H0415 H. Ovarian Tumor, II, OV5232 Ovarian Tumor, OV5232 Ovary disease pCMVSport 2.0 H0416 Human Neutrophils, Human Neutrophil - Blood Cell Line pBluescript Activated, re-excision Activated H0417 Human Pituitary, subtracted VIII Human Pituitary pBluescript H0418 Human Pituitary, subtracted VII Human Pituitary pBluescript H0419 Bone Cancer, re-excision Bone Cancer Uni-ZAP XR H0421 Human Bone Marrow, re-excision Bone Marrow pBluescript H0422 T-Cell PHA 16 hrs T-Cells Blood Cell Line pSport1 H0423 T-Cell PHA 24 hrs T-Cells Blood Cell Line pSport1 H0424 Human Pituitary, subt IX Human Pituitary pBluescript H0427 Human Adipose Human Adipose, left pSport1 hiplipoma H0428 Human Ovary Human Ovary Tumor Ovary pSport1 H0429 K562 + PMA (36 hrs), re-excision K562 Cell line cell line Cell Line ZAP Express H0431 H. Kidney Medulla, re-excision Kidney medulla Kidney pBluescript H0432 H. Kidney Pyramid Kidney pyramids Kidney pBluescript H0433 Human Umbilical Vein Endothelial cells, frac HUVE Cells Umbilical Cell Line pBluescript B, re-excision vein H0434 Human Brain, striatum, re-excision Human Brain, Striatum pBluescript H0435 Ovarian Tumor Oct. 3, 1995 Ovarian Tumor, OV350721 Ovary pCMVSport 2.0 H0436 Resting T-Cell Library, II T-Cells Blood Cell Line pSport1 H0437 H Umbilical Vein Endothelial Cells, frac A, HUVE Cells Umbilical Cell Line Lambda ZAP II re-excision vein H0438 H. Whole Brain #2, re-excision Human Whole Brain #2 ZAP Express H0439 Human Eosinophils Eosinophils pBluescript H0440 FGF enriched mixed library Mixed libraries pCMVSport 1 H0441 H. Kidney Cortex, subtracted Kidney cortex Kidney pBluescript H0442 H. Striatum Depression, subt II Human Brain, Striatum Brain pBluescript Depression H0443 H. Adipose, subtracted Human Adipose, left pSport1 hiplipoma H0444 Spleen metastic melanoma Spleen, Metastic Spleen disease pSport1 malignant melanoma H0445 Spleen, Chronic Human Spleen, CLL Spleen disease pSport1 lymphocytic leukemia H0447 Salivary gland, re-excision Human Salivary Gland Salivary gland Uni-ZAP XR H0448 Salivary gland, subtracted Human Salivary Gland Salivary gland Lambda ZAP II H0449 CD34 + cell, I CD34 positive cells pSport1 H0450 CD34 + cells, II CD34 positive cells pCMVSport 2.0 H0453 H. Kidney Pyramid, subtracted Kidney Pyramids Kidney pBluescript H0455 H. Striatum Depression, subt Human Brain, Striatum Brain pBluescript Depression H0456 H Kidney Cortex, subtracted III Human Kidney Cortex pBluescript H0457 Human Eosinophils Human Eosinophils pSport 1 H0458 CD34 + cell, I, frac II CD34 positive cells pSport 1 H0459 CD34 + cells, II, FRACTION 2 CD34 positive cells pCMVSport 2.0 H0461 H. Kidney Medulla, subtracted Kidney medulla Kidney pBluescript H0462 H. Amygdala Depression, subtracted Brain pBluescript H0477 Human Tonsil, Lib 3 Human Tonsil Tonsil pSport1 H0478 Salivary Gland, Lib 2 Human Salivary Gland Salivary gland pSport1 H0479 Salivary Gland, Lib 3 Human Salivary Gland Salivary gland pSport1 H0480 L8 cell line L8 cell line ZAP Express H0483 Breast Cancer cell line, Breast Cancer Cell pSport1 MDA 36 line, MDA 36 H0484 Breast Cancer Cell line, Breast Cancer Cell line, pSport1 angiogenic Angiogenic, 36T3 H0485 Hodgkin''s Lymphoma I Hodgkin''s Lymphoma I disease pCMVSport 2.0 H0486 Hodgkin''s Lymphoma II Hodgkin''s Lymphoma II disease pCMVSport 2.0 H0487 Human Tonsils, lib I Human Tonsils pCMVSport 2.0 H0488 Human Tonsils, Lib 2 Human Tonsils pCMVSport 2.0 H0489 Crohn''s Disease Ileum Intestine disease pSport1 H0490 Hl-60, untreated, subtracted Human HL-60 Cells, Blood Cell Line Uni-ZAP XR unstimulated H0491 HL-60, PMA 4 H, subtracted HL-60 Cells, PMA Blood Cell Line Uni-ZAP XR stimulated 4 H H0492 HL-60, RA 4 h, Subtracted HL-60 Cells, RA Blood Cell Line Uni-ZAP XR stimulated for 4 H H0493 HL-60, PMA 1 d, subtracted HL-60 Cells, PMA Blood Cell Line Uni-ZAP XR stimulated for 1 day H0494 Keratinocyte Keratinocyte pCMVSport 2.0 H0497 HEL cell line HEL cell line HEL 92.1.7 pSport1 H0505 Human Astrocyte Human Astrocyte pSport1 H0506 Ulcerative Colitis Colon Colon pSport1 H0509 Liver, Hepatoma Human Liver, Liver disease pCMVSport 3.0 Hepatoma, patient 8 H0510 Human Liver, normal Human Liver, Liver pCMVSport 3.0 normal, Patient # 8 H0512 Keratinocyte, lib 3 Keratinocyte pCMVSport 2.0 H0517 Nasal polyps Nasal polyps pCMVSport 2.0 H0518 pBMC stimulated w/poly I/C pBMC stimulated pCMVSport 3.0 with poly I/C H0519 NTERA2, control NTERA2, Teratocarcinoma pCMVSport 3.0 cell line H0520 NTERA2 + retinoic acid, NTERA2, Teratocarcinoma pSport1 14 days cell line H0521 Primary Dendritic Cells, lib 1 Primary Dendritic cells pCMVSport 3.0 H0522 Primary Dendritic cells, frac 2 Primary Dendritic cells pCMVSport 3.0 H0523 Primary Dendritic cells, CapFinder2, frac 1 Primary Dendritic cells pSport1 H0524 Primary Dendritic Cells, CapFinder, frac 2 Primary Dendritic cells pSport1 H0525 PCR, pBMC I/C treated pBMC stimulated with poly PCRII I/C H0528 Poly[I]/Poly[C] Normal Lung Fibroblasts Poly[I]/Poly[C] Normal pCMVSport 3.0 Lung Fibroblasts H0529 Myoloid Progenitor Cell Line TF-1 Cell Line; Myoloid pCMVSport 3.0 progenitor cell line H0530 Human Dermal Endothelial Cells, untreated Human Dermal Endothelial pSport1 Cells; untreated H0535 Human ovary tumor cell OV350721 Ovarian Tumor, OV350721 Ovary disease pSport1 H0537 H. Primary Dendritic Cells, lib 3 Primary Dendritic cells pCMVSport 2.0 H0538 Merkel Cells Merkel cells Lymph node pSport1 H0539 Pancreas Islet Cell Tumor Pancreas Islet Cell Tumour Pancreas disease pSport1 H0540 Skin, burned Skin, leg burned Skin Psport 1 H0542 T Cell helper I Helper T cell pCMVSport 3.0 H0543 T cell helper II Helper T cell pCMVSport 3.0 H0544 Human endometrial Human Endometrial pCMVSport 3.0 stromal cells stromal cells H0545 Human endometrial Human endometrial pCMVSport 3.0 stromal cells-treated with stromal cells-treated progesterone with proge H0546 Human endometrial Human endometrial pCMVSport 3.0 stromal cells-treated with stromal cells-treated estradiol with estra H0547 NTERA2 teratocarcinoma cell line + NTERA2, Teratocarcinoma pSport1 retinoic acid (14 days) cell line H0549 H. Epididiymus, caput & corpus Human Epididiymus, caput Uni-ZAP XR and corpus H0550 H. Epididiymus, cauda Human Epididiymus, cauda Uni-ZAP XR H0551 Human Thymus Stromal Cells Human Thymus Stromal Cells pCMVSport 3.0 H0552 Signal trap, Femur Bone Marrow, pooled Femur Bone marrow, pooled Other from 8 male/female H0553 Human Placenta Human Placenta pCMVSport 3.0 H0555 Rejected Kidney, lib 4 Human Rejected Kidney Kidney disease pCMVSport 3.0 H0556 Activated T-cell(12 h)/ T-Cells Blood Cell Line Uni-ZAP XR Thiouridine-re-excision H0559 HL-60, PMA 4 H, re-excision HL-60 Cells, PMA Blood Cell Line Uni-ZAP XR stimulated 4 H H0560 KMH2 KMH2 pCMVSport 3.0 H0561 L428 L428 pCMVSport 3.0 H0562 Human Fetal Brain, normalized c5-11-26 Human Fetal Brain pCMVSport 2.0 H0563 Human Fetal Brain, normalized 50021F Human Fetal Brain pCMVSport 2.0 H0564 Human Fetal Brain, normalized C5001F Human Fetal Brain pCMVSport 2.0 H0565 HUman Fetal Brain, normalized 100024F Human Fetal Brain pCMVSport 2.0 H0566 Human Fetal Brain, normalized c50F Human Fetal Brain pCMVSport 2.0 H0567 Human Fetal Brain, normalized A5002F Human Fetal Brain pCMVSport 2.0 H0569 Human Fetal Brain, normalized CO Human Fetal Brain pCMVSport 2.0 H0570 Human Fetal Brain, normalized C500H Human Fetal Brain pCMVSport 2.0 H0571 Human Fetal Brain, normalized C500HE Human Fetal Brain pCMVSport 2.0 H0572 Human Fetal Brain, normalized AC5002 Human Fetal Brain pCMVSport 2.0 H0574 Hepatocellular Tumor; re-excision Hepatocellular Tumor Liver disease Lambda ZAP II H0575 Human Adult Pulmonary; re-excision Human Adult Pulmonary Lung Uni-ZAP XR H0576 Resting T-Cell; re-excision T-Cells Blood Cell Line Lambda ZAP II H0578 Human Fetal Thymus Fetal Thymus Thymus pSport1 H0579 Pericardium Pericardium Heart pSport1 H0580 Dendritic cells, pooled Pooled dendritic cells pCMVSport 3.0 H0581 Human Bone Marrow, treated Human Bone Marrow Bone Marrow pCMVSport 3.0 H0583 B Cell lymphoma B Cell Lymphoma B Cell disease pCMVSport 3.0 H0584 Activated T-cells, 24 hrs, re-excision Activated T-Cells Blood Cell Line Uni-ZAP XR H0585 Activated T-Cells, 12 hrs, re-excision Activated T-Cells Blood Cell Line Uni-ZAP XR H0586 Healing groin wound, 6.5 healing groin wound, 6.5 hours groin disease pCMVSport 3.0 hours post incision post incision - 2/ H0587 Healing groin wound; 7.5 Groin-Feb. 19, 1997 groin disease pCMVSport 3.0 hours post incision H0589 CD34 positive cells (cord blood), re-ex CD34 Positive Cells Cord Blood ZAP Express H0590 Human adult small Human Adult Small Small Int. Uni-ZAP XR intestine, re-excision Intestine H0591 Human T-cell lymphoma; re-excision T-Cell Lymphoma T-Cell disease Uni-ZAP XR H0592 Healing groin wound - zero hr post-incision HGS wound healing disease pCMVSport 3.0 (control) project; abdomen H0593 Olfactory epithelium; nasalcavity Olfactory epithelium pCMVSport 3.0 from roof of left nasal cacit H0594 Human Lung Cancer; re-excision Human Lung Cancer Lung disease Lambda ZAP II H0595 Stomach cancer (human); re-excision Stomach Cancer - disease Uni-ZAP XR 5383A (human) H0596 Human Colon Cancer; re-excision Human Colon Cancer Colon Lambda ZAP II H0597 Human Colon; re-excision Human Colon Lambda ZAP II H0598 Human Stomach; re-excision Human Stomach Stomach Uni-ZAP XR H0599 Human Adult Heart; re-excision Human Adult Heart Heart Uni-ZAP XR H0600 Healing Abdomen wound; Abdomen disease pCMVSport 3.0 70&90 min post incision H0601 Healing Abdomen Wound; 15 days post Abdomen disease pCMVSport 3.0 incision H0602 Healing Abdomen Wound; 21&29 days post Abdomen disease pCMVSport 3.0 incision H0604 Human Pituitary, re-excision Human Pituitary pBluescript H0606 Human Primary Breast Human Primary Breast disease Uni-ZAP XR Cancer; re-excision Breast Cancer H0607 H. Leukocytes, normalized cot 50A3 H. Leukocytes pCMVSport 1 H0608 H. Leukocytes, control H. Leukocytes pCMVSport 1 H0609 H. Leukocytes, normalized cot > 500A H. Leukocytes pCMVSport 1 H0610 H. Leukocytes, normalized cot 5A H. Leukocytes pCMVSport 1 H0611 H. Leukocytes, normalized cot 500 B H. Leukocytes pCMVSport 1 H0612 H. Leukocytes, normalized cot 50 B H. Leukocytes pCMVSport 1 H0613 H. Leukocytes, normalized cot 5B H. Leukocytes pCMVSport 1 H0614 H. Leukocytes, normalized cot 500 A H. Leukocytes pCMVSport 1 H0615 Human Ovarian Cancer Reexcision Ovarian Cancer Ovary disease Uni-ZAP XR H0616 Human Testes, Reexcision Human Testes Testis Uni-ZAP XR H0617 Human Primary Breast Human Primary Breast disease Uni-ZAP XR Cancer Reexcision Breast Cancer H0618 Human Adult Testes, Human Adult Testis Testis Uni-ZAP XR Large Inserts, Reexcision H0619 Fetal Heart Human Fetal Heart Heart Uni-ZAP XR H0620 Human Fetal Kidney; Reexcision Human Fetal Kidney Kidney Uni-ZAP XR H0622 Human Pancreas Tumor; Reexcision Human Pancreas Tumor Pancreas disease Uni-ZAP XR H0623 Human Umbilical Vein; Reexcision Human Umbilical Vein Umbilical Uni-ZAP XR Endothelial Cells vein H0624 12 Week Early Stage Twelve Week Old Embryo Uni-ZAP XR Human II; Reexcision Early Stage Human H0625 Ku 812F Basophils Line Ku 812F Basophils pSport1 H0626 Saos2 Cells; Untreated Saos2 Cell Line; Untreated pSport1 H0627 Saos2 Cells; Vitamin D3 Treated Saos2 Cell Line; pSport1 Vitamin D3 Treated H0628 Human Pre-Differentiated Human Pre-Differentiated Uni-ZAP XR Adipocytes Adipocytes H0629 Human Leukocyte, control #2 Human Normalized pCMVSport 1 leukocyte H0630 Human Leukocytes, normalized control #4 Human Normalized pCMVSport 1 leukocyte H0631 Saos2, Dexamethosome Treated Saos2 Cell Line; pSport1 Dexamethosome Treated H0632 Hepatocellular Tumor; re-excision Hepatocellular Tumor Liver Lambda ZAP II H0633 Lung Carcinoma A549 TNFalpha activated disease pSport1 TNFalpha activated A549--Lung Carcinoma H0634 Human Testes Tumor, re-excision Human Testes Tumor Testis disease Uni-ZAP XR H0635 Human Activated T-Cells, re-excision Activated T-Cells Blood Cell Line Uni-ZAP XR H0637 Dendritic Cells From CD34 Cells Dendritic cells from pSport 1 CD34 cells H0638 CD40 activated monocyte CD40 activated monocyte pSport1 dendridic cells dendridic cells H0639 Ficolled Human Stromal Cells, 5Fu treated Ficolled Human Stromal Other Cells, 5Fu treated H0640 Ficolled Human Stromal Cells, Untreated Ficolled Human Stromal Other Cells, Untreated H0641 LPS activated derived LPS activated monocyte pSport1 dendritic cells derived dendritic cells H0642 Hep G2 Cells, lambda library Hep G2 Cells Other H0643 Hep G2 Cells, PCR library Hep G2 Cells Other H0644 Human Placenta (re-excision) Human Placenta Placenta Uni-ZAP XR H0645 Fetal Heart, re-excision Human Fetal Heart Heart Uni-ZAP XR H0646 Lung, Cancer (4005313 A3): Metastatic pSport1 Invasive Poorly Differentiated squamous cell lung Lung Adenocarcinoma, carcinoma, poorly di H0647 Lung, Cancer (4005163 B7): Invasive poorly disease pSport1 Invasive, Poorly Diff. differentiated lung Adenocarcinoma, Metastatic adenocarcinoma H0648 Ovary, Cancer: (4004562 B6) Papillary Papillary Cstic neoplasm of disease pSport1 Serous Cystic Neoplasm, Low Malignant Pot low malignant potentia H0649 Lung, Normal: (4005313 B1) Normal Lung pSport1 H0650 B-Cells B-Cells pCMVSport 3.0 H0651 Ovary, Normal: (9805C040R) Normal Ovary pSport1 H0652 Lung, Normal: (4005313 B1) Normal Lung pSport1 H0653 Stromal Cells Stromal Cells pSport1 H0654 Lung, Cancer: (4005313 A3) Invasive Metastatic Squamous cell Other Poorly-differentiated Metastatic lung adenoc lung Carcinoma poorly dif H0656 B-cells (unstimulated) B-cells (unstimulated) pSport1 H0657 B-cells (stimulated) B-cells (stimulated) pSport1 H0658 Ovary, Cancer (9809C332): 9809C332- Poorly Ovary & disease pSport1 Poorly differentiated adenocarcinoma differentiate Fallopian Tubes H0659 Ovary, Cancer (15395A1F): Grade II Grade II Papillary Ovary disease pSport1 Papillary Carcinoma Carcinoma, Ovary H0660 Ovary, Cancer: (15799A1F) Poorly Poorly differentiated disease pSport1 differentiated carcinoma carcinoma, ovary H0661 Breast, Cancer: (4004943 A5) Breast cancer disease pSport1 H0662 Breast, Normal: (4005522B2) Normal Breast - Breast pSport1 #4005522(B2) H0663 Breast, Cancer: (4005522 A2) Breast Cancer - Breast disease pSport1 #4005522(A2) H0664 Breast, Cancer: (9806C012R) Breast Cancer Breast disease pSport1 H0665 Stromal cells 3.88 Stromal cells 3.88 pSport1 H0666 Ovary, Cancer: (4004332 A2) Ovarian Cancer, disease pSport1 Sample #4004332A2 H0667 Stromal cells(HBM3.18) Stromal cell(HBM 3.18) pSport1 H0668 stromal cell clone 2.5 stromal cell clone 2.5 pSport1 H0669 Breast, Cancer: (4005385 A2) Breast Cancer (4005385A2) Breast pSport1 H0670 Ovary, Cancer(4004650 A3): Ovarian Cancer - pSport1 Well-Differentiated 4004650A3 Micropapillary Serous Carcinoma H0671 Breast, Cancer: (9802C02OE) Breast Cancer- Sample # pSport1 9802C02OE H0672 Ovary, Cancer: (4004576 A8) Ovarian Cancer(4004576A8) Ovary pSport1 H0673 Human Prostate Cancer, Human Prostate Prostate Uni-ZAP XR Stage B2; re-excision Cancer, stage B2 H0674 Human Prostate Cancer, Human Prostate Prostate Uni-ZAP XR Stage C; re-excission Cancer, stage C H0675 Colon, Cancer: (9808C064R) Colon Cancer 9808C064R pCMVSport 3.0 H0676 Colon, Cancer: (9808C064R)-total RNA Colon Cancer 9808C064R pCMVSport 3.0 H0677 TNFR degenerate oligo B-Cells PCRII H0678 screened clones from placental library Placenta Placenta Other H0679 screened clones from Tonsil library Human Tonsils Other H0682 Serous Papillary serous papillary pCMVSport 3.0 Adenocarcinoma adenocarcinoma (9606G304SPA3B) H0683 Ovarian Serous Papillary Adenocarcinoma Serous papillary pCMVSport 3.0 adenocarcinoma, stage 3C (9804G01 H0684 Serous Papillary Adenocarcinoma Ovarian Cancer- Ovaries pCMVSport 3.0 9810G606 H0685 Adenocarcinoma of Adenocarcinoma of pCMVSport 3.0 Ovary, Human Cell Line, Ovary, Human Cell # OVCAR-3 Line, # OVCAR- H0686 Adenocarcinoma of Adenocarcinoma of Ovary, pCMVSport 3.0 Ovary, Human Cell Line Human Cell Line, # SW-626 H0687 Human normal Human normal Ovary pCMVSport 3.0 ovary(#9610G215) ovary(#9610G215) H0688 Human Ovarian Cancer(#9807G017) Human Ovarian cancer pCMVSport 3.0 (#9807G017), mRNA from Maura Ru H0689 Ovarian Cancer Ovarian Cancer, #9806G019 pCMVSport 3.0 H0690 Ovarian Cancer, #9702G001 Ovarian Cancer, #9702G001 pCMVSport 3.0 H0691 Normal Ovary, #9710G208 normal ovary, #9710G208 pCMVSport 3.0 H0692 BLyS Receptor from Expression Cloning B Cell Lymphoma B Cell pCMVSport 3.0 H0693 Normal Prostate #ODQ3958EN Normal Prostate Tissue # pCMVSport 3.0 ODQ3958EN H0694 Prostate gland adenocarcinoma Prostate gland, prostate pCMVSport 3.0 adenocarcinoma, gland mod/diff, gleason H0695 mononucleocytes from patient mononucleocytes from patient pCMVSport 3.0 at Shady Grove Hospit N0006 Human Fetal Brain Human Fetal Brain N0007 Human Hippocampus Human Hippocampus N0009 Human Hippocampus, prescreened Human Hippocampus N0011 Human Brain Human Brain S0001 Brain frontal cortex Brain frontal cortex Brain Lambda ZAP II S0002 Monocyte activated Monocyte-activated blood Cell Line Uni-ZAP XR S0003 Human Osteoclastoma Osteoclastoma bone disease Uni-ZAP XR S0004 Prostate Prostate BPH Prostate Lambda ZAP II S0005 Heart Heart-left ventricle Heart pCDNA S0006 Neuroblastoma Human Neural Blastoma disease pCDNA S0007 Early Stage Human Brain Human Fetal Brain Uni-ZAP XR S0010 Human Amygdala Amygdala Uni-ZAP XR S0011 STROMAL -OSTEOCLASTOMA Osteoclastoma bone disease Uni-ZAP XR S0013 Prostate Prostate prostate Uni-ZAP XR S0014 Kidney Cortex Kidney cortex Kidney Uni-ZAP XR S0015 Kidney medulla Kidney medulla Kidney Uni-ZAP XR S0016 Kidney Pyramids Kidney pyramids Kidney Uni-ZAP XR S0021 Whole brain Whole brain Brain ZAP Express S0022 Human Osteoclastoma Osteoclastoma Uni-ZAP XR Stromal Cells - unamplified Stromal Cells S0024 Human Kidney Medulla - unamplified Human Kidney Medulla S0025 Human Kidney Pyramids - unamplified Human Kidney Pyramids S0026 Stromal cell TF274 stromal cell Bone marrow Cell Line Uni-ZAP XR S0027 Smooth muscle, serum treated Smooth muscle Pulmanary Cell Line Uni-ZAP XR artery S0028 Smooth muscle, control Smooth muscle Pulmanary Cell Line Uni-ZAP XR artery S0029 brain stem Brain stem brain Uni-ZAP XR S0030 Brain pons Brain Pons Brain Uni-ZAP XR S0031 Spinal cord Spinal cord spinal cord Uni-ZAP XR S0032 Smooth muscle-ILb induced Smooth muscle Pulmanary Cell Line Uni-ZAP XR artery S0035 Brain medulla oblongata Brain medulla oblongata Brain Uni-ZAP XR S0036 Human Substantia Nigra Human Substantia Nigra Uni-ZAP XR S0037 Smooth muscle, IL1b induced Smooth muscle Pulmanary Cell Line Uni-ZAP XR artery S0038 Human Whole Brain #2 - Oligo dT > 1.5 Kb Human Whole Brain #2 ZAP Express S0039 Hypothalamus Hypothalamus Brain Uni-ZAP XR S0040 Adipocytes Human Adipocytes Uni-ZAP XR from Osteoclastoma S0041 Thalamus Human Thalamus Uni-ZAP XR S0042 Testes Human Testes ZAP Express S0044 Prostate BPH prostate BPH Prostate disease Uni-ZAP XR S0045 Endothelial cells-control Endothelial cell endothelial Cell Line Uni-ZAP XR cell-lung S0046 Endothelial-induced Endothelial cell endothelial Cell Line Uni-ZAP XR cell-lung S0048 Human Hypothalamus, Alzheimer''s Human Hypothalamus, disease Uni-ZAP XR Alzheimer''s S0049 Human Brain, Striatum Human Brain, Striatum Uni-ZAP XR S0050 Human Frontal Cortex, Human Frontal Cortex, disease Uni-ZAP XR Schizophrenia Schizophrenia S0051 Human Hypothalmus, Human Hypothalamus, disease Uni-ZAP XR Schizophrenia Schizophrenia S0052 neutrophils control human neutrophils blood Cell Line Uni-ZAP XR S0053 Neutrophils IL-1 and LPS induced human neutrophil induced blood Cell Line Uni-ZAP XR S0106 STRIATUM DEPRESSION BRAIN disease Uni-ZAP XR S0110 Brain Amygdala Depression Brain disease Uni-ZAP XR S0112 Hypothalamus Brain Uni-ZAP XR S0114 Anergic T-cell Anergic T-cell Cell Line Uni-ZAP XR S0116 Bone marrow Bone marrow Bone marrow Uni-ZAP XR S0118 Smooth muscle control 2 Smooth muscle Pulmanary Cell Line Uni-ZAP XR artery S0122 Osteoclastoma-normalized A Osteoclastoma bone disease pBluescript S0124 Smooth muscle-edited A Smooth muscle Pulmanary Cell Line Uni-ZAP XR artery S0126 Osteoblasts Osteoblasts Knee Cell Line Uni-ZAP XR S0132 Epithelial-TNFa and INF induced Airway Epithelial Uni-ZAP XR S0134 Apoptotic T-cell apoptotic cells Cell Line Uni-ZAP XR S0136 PERM TF274 stromal cell Bone marrow Cell Line Lambda ZAP II S0140 eosinophil-IL5 induced eosinophil lung Cell Line Uni-ZAP XR S0142 Macrophage-oxLDL macrophage- blood Cell Line Uni-ZAP XR oxidized LDL treated S0144 Macrophage (GM-CSF treated) Macrophage (GM-CSF treated) Uni-ZAP XR S0146 prostate-edited prostate BPH Prostate Uni-ZAP XR S0148 Normal Prostate Prostate prostate Uni-ZAP XR S0150 LNCAP prostate cell line LNCAP Cell Line Prostate Cell Line Uni-ZAP XR S0152 PC3 Prostate cell line PC3 prostate cell line Uni-ZAP XR S0168 Prostate/LNCAP, subtraction I PC3 prostate cell line pBluescript S0174 Prostate-BPH subtracted II Human Prostate BPH pBluescript S0176 Prostate, normal, subtraction I Prostate prostate Uni-ZAP XR S0180 Bone Marrow Stroma, TNF&LPS ind Bone Marrow Stroma, TNF disease Uni-ZAP XR & LPS induced S0182 Human B Cell 8866 Human B- Cell 8866 Uni-ZAP XR S0184 7TM Receptor enriched, lib II PBLS, 7TM receptor enriched Other S0188 Prostate, BPH, Lib 2 Human Prostate BPH disease pSport1 S0190 Prostate BPH, Lib 2, subtracted Human Prostate BPH pSport1 S0192 Synovial Fibroblasts (control) Synovial Fibroblasts pSport1 S0194 Synovial hypoxia Synovial Fibroblasts pSport1 S0196 Synovial IL-1/TNF stimulated Synovial Fibroblasts pSport1 S0202 7TM-pbdd PBLS, 7TM receptor enriched PCRII S0206 Smooth Muscle- HASTE normalized Smooth muscle Pulmanary Cell Line pBluescript artery S0208 Messangial cell, frac 1 Messangial cell pSport1 S0210 Messangial cell, frac 2 Messangial cell pSport1 S0212 Bone Marrow Stromal Cell, untreated Bone Marrow Stromal pSport1 Cell, untreated S0214 Human Osteoclastoma, re-excision Osteoclastoma bone disease Uni-ZAP XR S0216 Neutrophils IL-1 and LPS induced human neutrophil induced blood Cell Line Uni-ZAP XR S0218 Apoptotic T-cell, re-excision apoptotic cells Cell Line Uni-ZAP XR S0220 H. hypothalamus, frac A; re-excision Hypothalamus Brain ZAP Express S0222 H. Frontal cortex; H. Brain, Frontal Brain disease Uni-ZAP XR epileptic, re-excision Cortex, Epileptic S0242 Synovial Fibroblasts (Il1/TNF), subt Synovial Fibroblasts pSport1 S0250 Human Osteoblasts II Human Osteoblasts Femur disease pCMVSport 2.0 S0252 7TM-PIMIX PBLS, 7TM receptor enriched PCRII S0256 7TM-PHMIX PBLS, 7TM receptor enriched PCRII S0260 Spinal Cord, re-excision Spinal cord spinal cord Uni-ZAP XR S0264 PPMIX PPMIX (Human Pituitary) Pituitary PCRII S0266 PLMIX PLMIX (Human Lung) Lung PCRII S0268 PRMIX PRMIX (Human Prostate) prostate PCRII S0270 PTMIX PTMIX (Human Thymus) Thymus PCRII S0276 Synovial hypoxia-RSF subtracted Synovial fobroblasts Synovial pSport1 (rheumatoid) tissue S0278 H Macrophage (GM-CSF Macrophage (GM- Uni-ZAP XR treated), re-excision CSF treated) S0280 Human Adipose Tissue, re-excision Human Adipose Tissue Uni-ZAP XR S0282 Brain Frontal Cortex, re-excision Brain frontal cortex Brain Lambda ZAP II S0292 Osteoarthritis (OA-4) Human Osteoarthritic Bone disease pSport1 Cartilage S0294 Larynx tumor Larynx tumor Larynx, vocal disease pSport1 cord S0296 Normal lung Normal lung Lung pSport1 S0298 Bone marrow stroma, treated Bone marrow stroma, treatedSB Bone marrow pSport1 S0300 Frontal lobe, dementia; re-excision Frontal Lobe Brain Uni-ZAP XR dementia/Alzheimer''s S0306 Larynx normal #10 261-273 Larynx normal pSport1 S0308 Spleen/normal Spleen normal pSport1 S0310 Normal trachea Normal trachea pSport1 S0312 Human osteoarthritic; fraction II Human osteoarthritic disease pSport1 cartilage S0314 Human osteoarthritis; fraction I Human osteoarthritic disease pSport1 cartilage S0316 Human Normal Cartilage, Fraction I Human Normal Cartilage pSport1 S0318 Human Normal Cartilage Fraction II Human Normal Cartilage pSport1 S0320 Human Larynx Larynx Epiglottis pSport1 S0322 Siebben Polyposis Siebben Polyposis pSport1 S0324 Human Brain Brain Cerebellum pSport1 S0328 Palate carcinoma Palate carcinoma Uvula disease pSport1 S0330 Palate normal Palate normal Uvula pSport1 S0332 Pharynx carcinoma Pharynx carcinoma Hypopharynx pSport1 S0334 Human Normal Cartilage Fraction III Human Normal Cartilage pSport1 S0336 Human Normal Cartilage Fraction IV Human Normal Cartilage pSport1 S0338 Human Osteoarthritic Human osteoarthritic disease pSport1 Cartilage Fraction III cartilage S0340 Human Osteoarthritic Cartilage Fraction IV Human osteoarthritic disease pSport1 cartilage S0342 Adipocytes; re-excision Human Adipocytes from Uni-ZAP XR Osteoclastoma S0344 Macrophage-oxLDL; re-excision macrophage- blood Cell Line Uni-ZAP XR oxidized LDL treated S0346 Human Amygdala; re-excision Amygdala Uni-ZAP XR S0348 Cheek Carcinoma Cheek Carcinoma disease pSport1 S0350 Pharynx Carcinoma Pharynx carcinoma Hypopharynx disease pSport1 S0352 Larynx Carcinoma Larynx carcinoma disease pSport1 S0354 Colon Normal II Colon Normal Colon pSport1 S0356 Colon Carcinoma Colon Carcinoma Colon disease pSport1 S0358 Colon Normal III Colon Normal Colon pSport1 S0360 Colon Tumor II Colon Tumor Colon disease pSport1 S0362 Human Gastrocnemius Gastrocnemius muscle pSport1 S0364 Human Quadriceps Quadriceps muscle pSport1 S0366 Human Soleus Soleus Muscle pSport1 S0368 Human Pancreatic Langerhans Islets of Langerhans pSport1 S0370 Larynx carcinoma II Larynx carcinoma disease pSport1 S0372 Larynx carcinoma III Larynx carcinoma disease pSport1 S0374 Normal colon Normal colon pSport1 S0376 Colon Tumor Colon Tumor disease pSport1 S0378 Pancreas normal PCA4 No Pancreas Normal PCA4 No pSport1 S0380 Pancreas Tumor PCA4 Tu Pancreas Tumor PCA4 Tu disease pSport1 S0382 Larynx carcinoma IV Larynx carcinoma disease pSport1 S0384 Tongue carcinoma Tongue carcinoma disease pSport1 S0386 Human Whole Brain, re-excision Whole brain Brain ZAP Express S0388 Human Hypothalamus, Human Hypothalamus, disease Uni-ZAP XR schizophrenia, re-excision Schizophrenia S0390 Smooth muscle, control; re-excision Smooth muscle Pulmanary Cell Line Uni-ZAP XR artery S0392 Salivary Gland Salivary gland; normal pSport1 S0394 Stomach; normal Stomach; normal pSport1 S0396 Uterus; normal Uterus; normal pSport1 S0398 Testis; normal Testis; normal pSport1 S0400 Brain; normal Brain; normal pSport1 S0402 Adrenal Gland, normal Adrenal gland; normal pSport1 S0404 Rectum normal Rectum, normal pSport1 S0406 Rectum tumour Rectum tumour pSport1 S0408 Colon, normal Colon, normal pSport1 S0410 Colon, tumour Colon, tumour pSport1 S0412 Temporal cortex-Alzheizmer; subtracted Temporal cortex, alzheimer disease Other S0414 Hippocampus, Alzheimer Subtracted Hippocampus, Alzheimer Other Subtracted S0418 CHME Cell Line; treated 5 hrs CHME Cell Line; treated pCMVSport 3.0 S0420 CHME Cell Line, untreated CHME Cell line, untreatetd pSport1 S0422 Mo7e Cell Line GM-CSF Mo7e Cell Line pCMVSport 3.0 treated (1 ng/ml) GM-CSF treated (1 ng/ml) S0424 TF-1 Cell Line GM-CSF Treated TF-1 Cell Line pSport1 GM-CSF Treated S0426 Monocyte activated; re-excision Monocyte-activated blood Cell Line Uni-Zap XR S0428 Neutrophils control; re-excision human neutrophils blood Cell Line Uni-ZAP XR S0430 Aryepiglottis Normal Aryepiglottis Normal pSport1 S0432 Sinus piniformis Tumour Sinus piniformis Tumour pSport1 S0434 Stomach Normal Stomach Normal disease pSport1 S0436 Stomach Tumour Stomach Tumour disease pSport1 S0438 Liver Normal Met5No Liver Normal Met5No pSport1 S0440 Liver Tumour Met 5 Tu Liver Tumour pSport1 S0442 Colon Normal Colon Normal pSport1 S0444 Colon Tumor Colon Tumour disease pSport1 S0446 Tongue Tumour Tongue Tumour pSport1 S0448 Larynx Normal Larynx Normal pSport1 S0450 Larynx Tumour Larynx Tumour pSport1 S0452 Thymus Thymus pSport1 S0454 Placenta Placenta Placenta pSport1 S0456 Tongue Normal Tongue Normal pSport1 S0458 Thyroid Normal (SDCA2 No) Thyroid normal pSport1 S0460 Thyroid Tumour Thyroid Tumour pSport1 S0462 Thyroid Thyroiditis Thyroid Thyroiditis pSport1 S0464 Larynx Normal Larynx Normal pSport1 S0466 Larynx Tumor Larynx Tumor disease pSport1 S0468 Ea.hy.926 cell line Ea.hy.926 cell line pSport1 S0470 Adenocarcinoma PYFD disease pSport1 S0472 Lung Mesothelium PYBT pSport1 S0474 Human blood platelets Platelets Blood Other platelets S0665 Human Amygdala; re-excission Amygdala Uni-ZAP XR S3012 Smooth Muscle Serum Smooth muscle Pulmanary Cell Line pBluescript Treated, Norm artery S3014 Smooth muscle, serum Smooth muscle Pulmanary Cell Line pBluescript induced, re-exc artery S6014 H. hypothalamus, frac A Hypothalamus Brain ZAP Express S6016 H. Frontal Cortex, Epileptic H. Brain, Frontal Cortex, Brain disease Uni-ZAP XR Epileptic S6022 H. Adipose Tissue Human Adipose Tissue Uni-ZAP XR S6024 Alzheimers, spongy change Alzheimer''s/Spongy change Brain disease Uni-ZAP XR S6026 Frontal Lobe, Dementia Frontal Lobe Brain Uni-ZAP XR dementia/Alzheimer''s S6028 Human Manic Depression Human Manic depression tissue Brain disease Uni-ZAP XR Tissue T0001 Human Brown Fat Brown Fat pBluescript SK- T0002 Activated T-cells Activated T-Cell, Blood Cell Line pBluescript SK- PBL fraction T0003 Human Fetal Lung Human Fetal Lung pBluescript SK- T0004 Human White Fat Human White Fat pBluescript SK- T0006 Human Pineal Gland Human Pinneal Gland pBluescript SK- T0007 Colon Epithelium Colon Epithelium pBluescriptISK- T0008 Colorectal Tumor Colorectal Tumor disease pBluescript SK T0010 Human Infant Brain Human Infant Brain Other T0023 Human Pancreatic Carcinoma Human Pancreatic Carcinoma disease pBluescript SK- T0039 HSA 172 Cells Human HSA172 cell line pBluescript SK- T0040 HSC172 cells SA172 Cells pBluescript SK- T0041 Jurkat T-cell G1 phase Jurkat T-cell pBluescript SK- T0042 Jurkat T Cell, S phase Jurkat T-Cell Line pBluescript SK- T0047 T lymphocytes >70 T lymphocytes >70 pBluescript SK- T0048 Human Aortic Endothelium Human Aortic Endothilium pBluescript SK- T0049 Aorta endothelial cells + TNF-a Aorta endothelial cells pBluescript SK- T0060 Human White Adipose Human White Fat pBluescript SK- T0067 Human Thyroid Human Thyroid pBluescript SK- T0068 Normal Ovary, Premenopausal Normal Ovary, Premenopausal pBluescript SK- T0069 Human Uterus, normal Human Uterus, normal pBluescript SK- T0070 Human Adrenal Gland Human Adrenal Gland pBluescript SK- T0071 Human Bone Marrow Human Bone Marrow pBluescript SK- T0078 Human Liver, normal adult Human Liver, normal Adult pBluescript SK- T0079 Human Kidney, normal Adult Human Kidney, normal Adult pBluescript SK- T0082 Human Adult Retina Human Adult Retina pBluescript SK- T0090 Liver, normal pBluescript SK- T0091 Liver, hepatocellular carcinoma pBluescript SK- T0104 HCC cell line metastisis to liver pBluescript SK- T0109 Human (HCC) cell line liver (mouse) pBluescript SK- metastasis, remake T0110 Human colon carcinoma (HCC) cell line, pBluescript SK- remake T0112 Human (Caco-2) cell line, adenocarcinoma, pBluescript SK- colon T0114 Human (Caco-2) cell line, pBluescript SK- adenocarcinoma, colon, remake T0115 Human Colon Carcinoma (HCC) cell line pBluescript SK- L0002 Atrium cDNA library Human heart L0004 ClonTech HL 1065a L0005 Clontech human aorta polyA + mRNA (#6572) L0009 EST from 8p21.3-p22 L0012 HDMEC cDNA library L0015 Human L0021 Human adult (K. Okubo) L0022 Human adult lung 3″ directed MboI cDNA L0024 Human brain ARSanders L0032 Human chromosome 12p cDNAs L0034 Human chromosome 14 L0038 Human chromosome 6 L0040 Human colon mucosa L0041 Human epidermal keratinocyte L0045 Human keratinocyte differential display (B. Lin) L0051 Human mRNA (Tripodis and Ragoussis) L0052 Human normalized K562-cDNA L0055 Human promyelocyte L0060 Human thymus NSTH II L0065 Liver HepG2 cell line. L0070 Selected chromosome 21 cDNA library L0096 Subtracted human retina L0097 Subtracted human retinal pigment epithelium (RPE) L0103 DKFZphamy1 amygdala L0105 Human aorta polyA+ (TFujiwara) aorta L0109 Human brain cDNA brain L0118 Human fetal brain S. Meier-Ewert brain L0126 Human fibroblast cDNA fibroblast L0138 Human normal gingiva normal gingiva L0141 Human pancreatic islet cell pancreatic islet L0142 Human placenta cDNA placenta (TFujiwara) L0143 Human placenta polyA+ (TFujiwara) placenta L0146 Human fovea cDNA retinal fovea L0149 DKFZphsnu1 subthalamic nucleus L0151 Human testis (C. De Smet) testis L0157 Human fetal brain (TFujiwara) brain L0158 Human fetal brain QBoqin brain L0162 Human brain frontal cortex frontal cortex brain L0163 Human heart cDNA (YNakamura) heart L0171 Human lung adenocarcinoma A549 lung adenocarcinoma A549 L0175 Human retina cell line ARPE-19 retina ARPE-19 L0177 Human newborn melanocytes (T. Vogt) Clonetics Corp. (San Diego, CA) strain #68 and 2486 L0182 Human HeLa (Y. Wang) HeLa L0186 Human salivary gland cell line HSG salivary gland HSG L0194 Human pancreatic cancer pancreatic cancer Patu cell line Patu 8988t 8988t L0351 Infant brain, Bento Soares BA, M13- derived L0352 Normalized infant brain, Bento Soares BA, M13-derived L0353 21q Placenta, F. Tassone and K. Gardiner Bluescript L0355 P, Human foetal Brain Whole tissue Bluescript L0356 S, Human foetal Adrenals tissue Bluescript L0361 Stratagene ovary (#937217) ovary Bluescript SK- L0362 Stratagene ovarian cancer (#937219) Bluescript SK- L0363 NCI_CGAP_GC2 germ cell tumor Bluescript SK- L0364 NCI_CGAP_GC5 germ cell tumor Bluescript SK- L0365 NCI_CGAP_Phe1 pheochromocytoma Bluescript SK- L0366 Stratagene schizo brain S11 schizophrenic brain Bluescript SK- S-11 frontal lobe L0367 NCI_CGAP_Sch1 Schwannoma tumor Bluescript SK- L0368 NCI_CGAP_SS1 synovial sarcoma Bluescript SK- L0369 NCI_CGAP_AA1 adrenal adenoma adrenal gland Bluescript SK- L0370 Johnston frontal cortex pooled frontal lobe brain Bluescript SK- L0371 NCI_CGAP_Br3 breast tumor breast Bluescript SK- L0372 NCI_CGAP_Co12 colon tumor colon Bluescript SK- L0373 NCI_CGAP_Co11 tumor colon Bluescript SK- L0374 NCI_CGAP_Co2 tumor colon Bluescript SK- L0375 NCI_CGAP_Kid6 kidney tumor kidney Bluescript SK- L0376 NCI_CGAP_Lar1 larynx larynx Bluescript SK- L0377 NCI_CGAP_HN2 squamous cell carcinoma larynx Bluescript SK- from vocal cord L0378 NCI_CGAP_Lu1 lung tumor lung Bluescript SK- L0379 NCI_CGAP_Lym3 lymphoma lymph node Bluescript SK- L0380 NCI_CGAP_HN1 squamous cell carcinoma lymph node Bluescript SK- L0381 NCI_CGAP_HN4 squamous cell carcinoma pharynx Bluescript SK- L0382 NCI_CGAP_Pr25 epithelium (cell line) prostate Bluescript SK- L0383 NCI_CGAP_Pr24 invasive tumor (cell line) prostate Bluescript SK- L0384 NCI_CGAP_Pr23 prostate tumor prostate Bluescript SK- L0385 NCI_CGAP_Gas1 gastric tumor stomach Bluescript SK- L0386 NCI_CGAP_HN3 squamous cell carcinoma tongue Bluescript SK- from base of tongue L0387 NCI_CGAP_GCB0 germinal center B-cells tonsil Bluescript SK- L0388 NCI_CGAP_HN6 normal gingiva (cell line Bluescript SK- from immortalized kerati L0389 NCI_CGAP_HN5 normal gingiva (cell line Bluescript SK- from primary keratinocyt L0393 B, Human Liver tissue gt11 L0394 H, Human adult Brain Cortex tissue gt11 L0404 b4HB3MA Cot109 + 103 + 85 - Bio Lafmid A L0405 b4HB3MA Cot109 + 103 - Bio Lafmid A L0411 1-NIB Lafmid BA L0414 b4HB3MA Lafmid BA L0415 b4HB3MA Cot8-HAP-Ft Lafmid BA L0416 b4HB3MA-Cot0.38-HAP-B Lafmid BA L0417 b4HB3MA-Cot0.38-HAP-Ft-6 Lafmid BA L0422 b4HB3MA-Cot12-HAP-B Lafmid BA L0424 b4HB3MA-Cot14.5 Lafmid BA L0425 b4HB3MA-Cot18-Bio Lafmid BA L0426 b4HB3MA-Cot51.5-HAP-Ft Lafmid BA L0427 b4HB3MA-FT20%-Biotin Lafmid BA L0428 Cot1374Ft-4HB3MA Lafmid BA L0430 Cot250Ft-b4HB3MA Lafmid BA L0434 Infant brain library of Dr. M. Soares lafmid BA L0435 Infant brain, LLNL array of lafmid BA Dr. M. Soares 1NIB L0437 N-b4HB3MA-Cot109 Lafmid BA L0438 normalized infant brain cDNA total brain brain lafmid BA L0439 Soares infant brain 1NIB whole brain Lafmid BA L0441 2HB3MK Lafmid BK L0442 4HB3MK Lafmid BK L0443 b4HB3MK Lafmid BK L0446 N4HB3MK Lafmid BK L0447 NHB3MK Lafmid BK L0448 3HFLSK20 Lafmid K L0451 N3HFLSK20 Lafmid K L0453 BATM1 lambda gt10 L0454 Clontech adult human fat cell library lambda gt10 HL1108A L0455 Human retina cDNA randomly primed retina eye lambda gt10 sublibrary L0456 Human retina cDNA Tsp509I-cleaved retina eye lambda gt10 sublibrary L0457 multi-tissue normalized short-fragment multi-tissue pooled lambda gt10 L0459 Adult heart, Clontech Lambda gt11 L0460 Adult heart, Lambda gt11 Lambda gt11 L0462 WATM1 lambda gt11 L0463 fetal brain cDNA brain brain lambda gt11 L0465 TEST1, Human adult Testis tissue lambda nm1149 L0468 HE6W lambda zap L0469 T, Human adult Rhabdomyosarcoma cell-line Lambda Zap L0470 BL29 Burkitt''s lymphoma, Pascalis Sideras lambda ZAP 2 L0471 Human fetal heart, Lambda ZAP Express Lambda ZAP Express L0475 KG1-a Lambda Zap KG1-a Lambda Zap Express cDNA library Express (Stratagene) L0476 Fetal brain, Stratagene Lambda ZAP II L0477 HPLA CCLee placenta Lambda ZAP II L0480 Stratagene cat#937212 (1992) Lambda ZAP, pBluescript SK(-) L0481 CD34 + DIRECTIONAL Lambda ZAPII L0483 Human pancreatic islet Lambda ZAPII L0485 STRATAGENE Human skeletal muscle skeletal muscle leg muscle Lambda ZAPII cDNA library, cat. #936215. L0487 Human peripheral blood (Steve Elledge) whole peripheral blood Lambda-Yes L0492 Human Genomic pAMP L0493 NCI_CGAP_Ov26 papillary serous carcinoma ovary pAMP1 L0497 NCI_CGAP_HSC4 CD34+, CD38- from normal bone marrow pAMP1 bone marrow donor L0498 NCI_CGAP_HSC3 CD34+, T negative, patient bone marrow pAMP1 with chronic myelogenou L0499 NCI_CGAP_HSC2 stem cell 34+/38+ bone marrow pAMP1 L0500 NCI_CGAP_Brn20 oligodendroglioma brain pAMP1 L0501 NCI_CGAP_Brn21 oligodendroglioma brain pAMP1 L0502 NCI_CGAP_Br15 adenocarcinoma breast pAMP1 L0503 NCI_CGAP_Br17 adenocarcinoma breast pAMP1 L0504 NCI_CGAP_Br13 breast carcinoma in situ breast pAMP1 L0505 NCI_CGAP_Br12 invasive carcinoma breast pAMP1 L0506 NCI_CGAP_Br16 lobullar carcinoma in situ breast pAMP1 L0507 NCI_CGAP_Br14 normal epithelium breast pAMP1 L0508 NCI_CGAP_Lu25 bronchioalveolar carcinoma lung pAMP1 L0509 NCI_CGAP_Lu26 invasive adenocarcinoma lung pAMP1 L0510 NCI_CGAP_Ov33 borderline ovarian carcinoma ovary pAMP1 L0511 NCI_CGAP_Ov34 borderline ovarian carcinoma ovary pAMP1 L0512 NCI_CGAP_Ov36 borderline ovarian carcinoma ovary pAMP1 L0513 NCI_CGAP_Ov37 early stage papillary serous carcinoma ovary pAMP1 L0514 NCI_CGAP_Ov31 papillary serous carcinoma ovary pAMP1 L0515 NCI_CGAP_Ov32 papillary serous carcinoma ovary pAMP1 L0516 Chromosome 19p12-p13.1 exon pAMP10 L0517 NCI_CGAP_Pr1 pAMP10 L0518 NCI_CGAP_Pr2 pAMP10 L0519 NCI_CGAP_Pr3 pAMP10 L0520 NCI_CGAP_Alv1 alveolar rhabdomyosarcoma pAMP10 L0521 NCI_CGAP_Ew1 Ewing''s sarcoma pAMP10 L0522 NCI_CGAP_Kid1 kidney pAMP10 L0523 NCI_CGAP_Lip2 liposarcoma pAMP10 L0524 NCI_CGAP_Li1 liver pAMP10 L0525 NCI_CGAP_Li2 liver pAMP10 L0526 NCI_CGAP_Pr12 metastatic prostate pAMP10 bone lesion L0527 NCI_CGAP_Ov2 ovary pAMP10 L0528 NCI_CGAP_Pr5 prostate pAMP10 L0529 NCI_CGAP_Pr6 prostate pAMP10 L0530 NCI_CGAP_Pr8 prostate pAMP10 L0531 NCI_CGAP_Pr20 prostate metastasis, liver pAMP10 L0532 NCI_CGAP_Thy1 thyroid pAMP10 L0533 NCI_CGAP_HSC1 stem cells bone marrow pAMP10 L0534 Chromosome 7 Fetal Brain cDNA Library brain brain pAMP10 L0535 NCI_CGAP_Br5 infiltrating ductal carcinoma breast pAMP10 L0536 NCI_CGAP_Br4 normal ductal tissue breast pAMP10 L0537 NCI_CGAP_Ov6 normal cortical stroma ovary pAMP10 L0539 Chromosome 7 Placental cDNA Library placenta pAMP10 L0540 NCI_CGAP_Pr10 invasive prostate tumor prostate pAMP10 L0541 NCI_CGAP_Pr7 low-grade prostatic prostate pAMP10 neoplasia L0542 NCI_CGAP_Pr11 normal prostatic prostate pAMP10 epithelial cells L0543 NCI_CGAP_Pr9 normal prostatic prostate pAMP10 epithelial cells L0544 NCI_CGAP_Pr4 prostatic intraepithelial prostate pAMP10 neoplasia - high grade L0545 NCI_CGAP_Pr4.1 prostatic intraepithelial prostate pAMP10 neoplasia - high grade L0546 NCI_CGAP_Pr18 stroma prostate pAMP10 L0547 NCI_CGAP_Pr16 tumor prostate pAMP10 L0548 Chromosome 7 Thymus cDNA Library thymus thymus pAMP10 L0549 NCI_CGAP_HN10 carcinoma in situ from pAMP10 retromolar trigone L0550 NCI_CGAP_HN9 normal squamous epithelium pAMP10 from retromolar trigone L0551 NCI_CGAP_HN7 normal squamous pAMP10 epithelium, floor of mouth L0552 NCI_CGAP_HN8 well-differentiated invasive pAMP10 carcinoma, floor of m L0553 NCI_CGAP_Co22 colonic adenocarcinoma colon pAMP10 L0554 NCI_CGAP_Li8 liver pAMP10 L0555 NCI_CGAP_Lu34 large cell carcinoma lung pAMP10 L0556 NCI_CGAP_Lu34.1 large cell carcinoma lung pAMP10 L0558 NCI_CGAP_Ov40 endometrioid ovarian ovary pAMP10 metastasis L0559 NCI_CGAP_Ov39 papillary serous ovary pAMP10 ovarian metastasis L0560 NCI_CGAP_HN12 moderate to poorly tongue pAMP10 differentiated invasive carcino L0561 NCI_CGAP_HN11 normal squamous epithelium tongue pAMP10 L0562 Chromosome 7 HeLa cDNA Library HeLa cell pAMP10 line; ATCC L0563 Human Bone Marrow Stromal Fibroblast bone marrow pBluescript L0564 Jia bone marrow stroma bone marrow stroma pBluescript L0565 Normal Human Bone Hip pBluescript Trabecular Bone Cells L0579 Human fetal brain QBoqin2 cerebrum and cerebellum pBluescript SK L0581 Stratagene liver (#937224) liver pBluescript SK L0583 Stratagene cDNA library Human fibroblast, pBluescript SK(+) cat#937212 L0584 Stratagene cDNA library Human heart, pBluescript SK(+) cat#936208 L0586 HTCDL1 pBluescript SK(-) L0587 Stratagene colon HT29 (#937221) pBluescript SK- L0588 Stratagene endothelial cell 937223 pBluescript SK- L0589 Stratagene fetal retina 937202 pBluescript SK- L0590 Stratagene fibroblast (#937212) pBluescript SK- L0591 Stratagene HeLa cell s3 937216 pBluescript SK- L0592 Stratagene hNT neuron (#937233) pBluescript SK- L0593 Stratagene neuroepithelium (#937231) pBluescript SK- L0594 Stratagene neuroepithelium pBluescript SK- NT2RAMI 937234 L0595 Stratagene NT2 neuronal neuroepithelial cells brain pBluescript SK- precursor 937230 L0596 Stratagene colon (#937204) colon pBluescript SK- L0597 Stratagene corneal stroma (#937222) cornea pBluescript SK- L0598 Morton Fetal Cochlea cochlea ear pBluescript SK- L0599 Stratagene lung (#937210) lung pBluescript SK- L0600 Weizmann Olfactory Epithelium olfactory epithelium nose pBluescript SK- L0601 Stratagene pancreas (#937208) pancreas pBluescript SK- L0602 Pancreatic Islet pancreatic islet pancreas pBluescript SK- L0603 Stratagene placenta (#937225) placenta pBluescript SK- L0604 Stratagene muscle 937209 muscle skeletal pBluescript SK- muscle L0605 Stratagene fetal spleen (#937205) fetal spleen spleen pBluescript SK- L0606 NCI_CGAP_Lym5 follicular lymphoma lymph node pBluescript SK- L0607 NCI_CGAP_Lym6 mantle cell lymphoma lymph node pBluescript SK- L0608 Stratagene lung carcinoma 937218 lung carcinoma lung NCI-H69 pBluescript SK- L0609 Schiller astrocytoma astrocytoma brain pBluescript SK- (Stratagene) L0611 Schiller meningioma meningioma brain pBluescript SK- (Stratagene) L0612 Schiller oligodendroglioma oligodendroglioma brain pBluescript SK- (Stratagene) L0615 22 week old human fetal liver cDNA library pBluescriptII SK(-) L0617 Chromosome 22 exon pBluescriptIIKS+ L0619 Chromosome 9 exon II pBluescriptIIKS+ L0622 HM1 pcDNAII (Invitrogen) L0623 HM3 pectoral muscle (after pcDNAII mastectomy) (Invitrogen) L0625 NCI_CGAP_AR1 bulk alveolar tumor pCMV-SPORT2 L0626 NCI_CGAP_GC1 bulk germ cell seminoma pCMV-SPORT2 L0627 NCI_CGAP_Co1 bulk tumor colon pCMV-SPORT2 L0628 NCI_CGAP_Ov1 ovary bulk tumor ovary pCMV-SPORT2 L0629 NCI_CGAP_Mel3 metastatic melanoma to bowel (skin pCMV-SPORT4 bowel primary) L0630 NCI_CGAP_CNS1 substantia nigra brain pCMV-SPORT4 L0631 NCI_CGAP_Br7 breast pCMV-SPORT4 L0632 NCI_CGAP_Li5 hepatic adenoma liver pCMV-SPORT4 L0633 NCI_CGAP_Lu6 small cell carcinoma lung pCMV-SPORT4 L0634 NCI_CGAP_Ov8 serous adenocarcinoma ovary pCMV-SPORT4 L0635 NCI_CGAP_PNS1 dorsal root ganglion peripheral pCMV-SPORT4 nervous system L0636 NCI_CGAP_Pit1 four pooled pituitary brain pCMV-SPORT6 adenomas L0637 NCI_CGAP_Brn53 three pooled meningiomas brain pCMV-SPORT6 L0638 NCI_CGAP_Brn35 tumor, 5 pooled (see brain pCMV-SPORT6 description) L0639 NCI_CGAP_Brn52 tumor, 5 pooled (see brain pCMV-SPORT6 description) L0640 NCI_CGAP_Br18 four pooled high-grade breast pCMV-SPORT6 tumors, including two prima L0641 NCI_CGAP_Co17 juvenile granulosa tumor colon pCMV-SPORT6 L0642 NCI_CGAP_Co18 moderately differentiated colon pCMV-SPORT6 adenocarcinoma L0643 NCI_CGAP_Co19 moderately differentiated colon pCMV-SPORT6 adenocarcinoma L0644 NCI_CGAP_Co20 moderately colon pCMV-SPORT6 differentiated adenocarcinoma L0645 NCI_CGAP_Co21 moderately colon pCMV-SPORT6 differentiated adenocarcinoma L0646 NCI_CGAP_Co14 moderately- colon pCMV-SPORT6 differentiated adenocarcinoma L0647 NCI_CGAP_Sar4 five pooled connective pCMV-SPORT6 sarcomas, including tissue myxoid liposarcoma L0648 NCI_CGAP_Eso2 squamous cell esophagus pCMV-SPORT6 carcinoma L0649 NCI_CGAP_GU1 2 pooled high-grade genitourinary pCMV-SPORT6 transitional cell tract tumors L0650 NCI_CGAP_Kid13 2 pooled Wilms'' kidney pCMV-SPORT6 tumors, one primary and one metast L0651 NCI_CGAP_Kid8 renal cell tumor kidney pCMV-SPORT6 L0652 NCI_CGAP_Lu27 four pooled poorly- lung pCMV-SPORT6 differentiated adenocarcinomas L0653 NCI_CGAP_Lu28 two pooled squamous lung pCMV-SPORT6 cell carcinomas L0654 NCI_CGAP_Lu31 lung, cell line pCMV-SPORT6 L0655 NCI_CGAP_Lym12 lymphoma, lymph node pCMV-SPORT6 follicular mixed small and large cell L0656 NCI_CGAP_Ov38 normal epithelium ovary pCMV-SPORT6 L0657 NCI_CGAP_Ov23 tumor, 5 pooled (see ovary pCMV-SPORT6 description) L0658 NCI_CGAP_Ov35 tumor, 5 pooled (see ovary pCMV-SPORT6 description) L0659 NCI_CGAP_Pan1 adenocarcinoma pancreas pCMV-SPORT6 L0661 NCI_CGAP_Mel15 malignant melanoma, skin pCMV-SPORT6 metastatic to lymph node L0662 NCI_CGAP_Gas4 poorly differentiated stomach pCMV-SPORT6 adenocarcinoma with signet r L0663 NCI_CGAP_Ut2 moderately-differentiated uterus pCMV-SPORT6 endometrial adenocarcino L0664 NCI_CGAP_Ut3 poorly-differentiated uterus pCMV-SPORT6 endometrial adenocarcinoma, L0665 NCI_CGAP_Ut4 serous papillary uterus pCMV-SPORT6 carcinoma, high grade, 2 pooled t L0666 NCI_CGAP_Ut1 well-differentiated uterus pCMV-SPORT6 endometrial adenocarcinoma, 7 L0667 NCI_CGAP_CML1 myeloid cells, 18 whole blood pCMV SPORT6 pooled CML cases, BCR/ABL rearra L0669 Human MCF7 cDNA subtracted with MDA- breast adenocarcinoma breast MCF7 pCR II MB-231 cDNA [Invitrogen] L0681 Stanley Frontal SN individual frontal lobe (see brain pCR2.1 description) (Invitrogen) L0682 Stanley Frontal MB pool 2 frontal lobe (see brain pCR2.1-TOPO description) (Invitrogen) L0683 Stanley Frontal NS pool 2 frontal lobe (see brain pCR2.1-TOPO description) (Invitrogen) L0684 Stanley Frontal SB pool 1 frontal lobe (see brain pCR2.1-TOPO description) (Invitrogen) L0685 Stanley Frontal SN pool 1 frontal lobe (see brain pCR2.1-TOPO description) (Invitrogen) L0686 Stanley Frontal SN pool 2 frontal lobe (see brain pCR2.1-TOPO description) (Invitrogen) L0687 Stanley Hippocampus NB pool 1 hippocampus (see brain pCR2.1-TOPO description) (Invitrogen) L0688 Stanley Hippocampus SB pool 1 hippocampus (see brain pCR2.1-TOPO description) (Invitrogen) L0689 Stanley Hippocampus SN pool 1 hippocampus (see brain pCR2.1-TOPO description) (Invitrogen) L0695 Human Glialblastoma Cell Brain BT-325 PCRII, Invitrogen L0697 Testis 1 PGEM 5zf(+) L0698 Testis 2 PGEM 5zf(+) L0700 Outward Alu-primed hncDNA library pGEM-3Z L0709 NIH_MGC_21 choriocarcinoma placenta pOTB7 L0710 NIH_MGC_7 small cell carcinoma lung MGC3 pOTB7 L0716 PMA-induced HL60 cell subtraction PMA- pSPORT 1 library induced HL60 human leukemic cell line L0717 Gessler Wilms tumor pSPORT1 L0718 Testis 5 pSPORT1 L0719 human embryo cDNA library Whole embryo pSPORT1 L0731 Soares_pregnant_uterus_(—) uterus pT7T3-Pac NbHPU L0738 Human colorectal cancer pT7T3D L0740 Soares melanocyte melanocyte pT7T3D 2NbHM (Pharmacia) with a modified polylinker L0741 Soares adult brain brain pT7T3D N2b4HB55Y (Pharmacia) with a modified polylinker L0742 Soares adult brain brain pT7T3D N2b5HB55Y (Pharmacia) with a modified polylinker L0743 Soares breast 2NbHBst breast pT7T3D (Pharmacia) with a modified polylinker L0744 Soares breast 3NbHBst breast pT7T3D (Pharmacia) with a modified polylinker L0745 Soares retina N2b4HR retina eye pT7T3D (Pharmacia) with a modified polylinker L0746 Soares retina N2b5HR retina eye pT7T3D (Pharmacia) with a modified polylinker L0747 Soares_fetal_heart_(—) heart pT7T3D NbHH19W (Pharmacia) with a modified polylinker L0748 Soares fetal liver spleen Liver and pT7T3D 1NFLS Spleen (Pharmacia) with a modified polylinker L0749 Soares_fetal_liver_(—) Liver and pT7T3D spleen_1NFLS_S1 Spleen (Pharmacia) with a modified polylinker L0750 Soares_fetal_lung_(—) lung pT7T3D NbHL19W (Pharmacia) with a modified polylinker L0751 Soares ovary tumor ovarian tumor ovary pT7T3D NbHOT (Pharmacia) with a modified polylinker L0752 Soares_parathyroid_(—) parathyroid tumor parathyroid pT7T3D tumor_NbHPA gland (Pharmacia) with a modified polylinker L0753 Soares_pineal_gland_(—) pineal gland pT7T3D N3HPG (Pharmacia) with a modified polylinker L0754 Soares placenta Nb2HP placenta pT7T3D (Pharmacia) with a modified polylinker L0755 Soares_placenta_(—) placenta pT7T3D 8to9weeks_2NbHP8to9W (Pharmacia) with a modified polylinker L0756 Soares_multiple_(—) multiple sclerosis pT7T3D sclerosis_2NbHMSP lesions (Pharmacia) with a modified polylinker V_TYPE L0757 Soares_senescent_(—) senescent fibroblast pT7T3D fibroblasts_NbHSF (Pharmacia) with a modified polylinker V_TYPE L0758 Soares_testis_NHT pT7T3D-Pac (Pharmacia) with a modified polylinker L0759 Soares_total_fetus_(—) pT7T3D-Pac Nb2HF8_9w (Pharmacia) with a modified polylinker L0760 Barstead aorta HPLRB3 aorta pT7T3D-Pac (Pharmacia) with a modified polylinker L0761 NCI_CGAP_CLL1 B-cell, chronic pT7T3D-Pac lymphotic leukemia (Pharmacia) with a modified polylinker L0762 NCI_CGAP_Br1.1 breast pT7T3D-Pac (Pharmacia) with a modified polylinker L0763 NCI_CGAP_Br2 breast pT7T3D-Pac (Pharmacia) with a modified polylinker L0764 NCI_CGAP_Co3 colon pT7T3D-Pac (Pharmacia) with a modified polylinker L0765 NCI_CGAP_Co4 colon pT7T3D-Pac (Pharmacia) with a modified polylinker L0766 NCI_CGAP_GCB1 germinal center B pT7T3D-Pac cell (Pharmacia) with a modified polylinker L0767 NCI_CGAP_GC3 pooled germ cell tumors pT7T3D-Pac (Pharmacia) with a modified polylinker L0768 NCI_CGAP_GC4 pooled germ cell pT7T3D-Pac tumors (Pharmacia) with a modified polylinker L0769 NCI_CGAP_Brn25 anaplastic brain pT7T3D-Pac oligodendroglioma (Pharmacia) with a modified polylinker L0770 NCI_CGAP_Brn23 glioblastoma brain pT7T3D-Pac (pooled) (Pharmacia) with a modified polylinker L0771 NCI_CGAP_Co8 adenocarcinoma colon pT7T3D-Pac (Pharmacia) with a modified polylinker L0772 NCI_CGAP_Co10 colon tumor RER+ colon pT7T3D-Pac (Pharmacia) with a modified polylinker L0773 NCI_CGAP_Co9 colon tumor RER+ colon pT7T3D-Pac (Pharmacia) with a modified polylinker L0774 NCI_CGAP_Kid3 kidney pT7T3D-Pac (Pharmacia) with a modified polylinker L0775 NCI_CGAP_Kid5 2 pooled tumors kidney pT7T3D-Pac (clear cell type) (Pharmacia) with a modified polylinker L0776 NCI_CGAP_Lu5 carcinoid lung pT7T3D-Pac (Pharmacia) with a modified polylinker L0777 Soares_NhHMPu_S1 Pooled human mixed (see pT7T3D-Pac melanocyte, fetal below) (Pharmacia) heart, and pregnant with a modified polylinker L0778 Barstead pancreas pancreas pT7T3D-Pac HPLRB1 (Pharmacia) with a modified polylinker L0779 Soares_NFL_T_GBC_S1 pooled pT7T3D-Pac (Pharmacia) with a modified polylinker L0780 Soares_NSF_F8_9W_(—) pooled pT7T3D-Pac OT_PA_P_S1 (Pharmacia) with a modified polylinker L0782 NCI_CGAP_Pr21 normal prostate prostate pT7T3D-Pac (Pharmacia) with a modified polylinker L0783 NCI_CGAP_Pr22 normal prostate prostate pT7T3D-Pac (Pharmacia) with a modified polylinker L0784 NCI_CGAP_Lei2 leiomyosarcoma soft tissue pT7T3D-Pac (Pharmacia) with a modified polylinker L0785 Barstead spleen HPLRB2 spleen pT7T3D-Pac (Pharmacia) with a modified polylinker L0786 Soares_NbHFB whole brain pT7T3D-Pac (Pharmacia) with a modified polylinker L0787 NCI_CGAP_Sub1 pT7T3D-Pac (Pharmacia) with a modified polylinker L0788 NCI_CGAP_Sub2 pT7T3D-Pac (Pharmacia) with a modified polylinker L0789 NCI_CGAP_Sub3 pT7T3D-Pac (Pharmacia) with a modified polylinker L0790 NCI_CGAP_Sub4 pT7T3D-Pac (Pharmacia) with a modified polylinker L0791 NCI_CGAP_Sub5 pT7T3D-Pac (Pharmacia) with a modified polylinker L0792 NCI_CGAP_Sub6 pT7T3D-Pac (Pharmacia) with a modified polylinker L0793 NCI_CGAP_Sub7 pT7T3D-Pac (Pharmacia) with a modified polylinker L0794 NCI_CGAP_GC6 pooled germ cell pT7T3D-Pac tumors (Pharmacia) with a modified polylinker L0796 NCI_CGAP_Brn50 medulloblastoma brain pT7T3D-Pac (Pharmacia) with a modified polylinker L0800 NCI_CGAP_Co16 colon tumor, RER+ colon pT7T3D-Pac (Pharmacia) with a modified polylinker L0803 NCI_CGAP_Kid11 kidney pT7T3D-Pac (Pharmacia) with a modified polylinker L0804 NCI_CGAP_Kid12 2 pooled tumors kidney pT7T3D-Pac (clear cell type) (Pharmacia) with a modified polylinker L0805 NCI_CGAP_Lu24 carcinoid lung pT7T3D-Pac (Pharmacia) with a modified polylinker L0806 NCI_CGAP_Lu19 squamous cell lung pT7T3D-Pac carcinoma, poorly (Pharmacia) differentiated (4 with a modified polylinker L0807 NCI_CGAP_Ov18 fibrotheoma ovary pT7T3D-Pac (Pharmacia) with a modified polylinker L0808 Barstead prostate BPH HPLRB4 1 prostate pT7T3D-Pac (Pharmacia) with a modified polylinker L0809 NCI_CGAP_Pr28 prostate pT7T3D-Pac (Pharmacia) with a modified polylinker L0811 BATM2 PTZ18 L0988 BT0387 breast puc 18 L1430 CT0225 colon puc 18 L1562 CN0027 colon_normal puc 18 L1819 HT0268 head_neck puc 18 L2138 ST0186 stomach puc 18 L2245 NEM subtracted human fetal kidney cDNA pUEX1 L2250 Human cerebral cortex cerebral cortex L2251 Human fetal lung Fetal lung L2252 Human placenta placenta L2255 GLC corresponding non pBluescript sk(-) cancerous liver tissue L2257 NIH_MGC_65 adenocarcinoma colon pCMV-SPORT6 L2258 NIH_MGC_67 retinoblastoma eye pCMV-SPORT6 L2259 NIH_MGC_68 large cell lung pCMV-SPORT6 carcinoma L2260 NIH_MGC_69 large cell carcinoma, lung pCMV-SPORT6 undifferentiated L2261 NIH_MGC_70 epithelioid pancreas pCMV-SPORT6 carcinoma L2262 NIH_MGC_72 melanotic melanoma skin pCMV-SPORT6 L2263 NIH_MGC_66 adenocarcinoma ovary pCMV-SPORT6 L2264 NIH_MGC_71 leiomyosarcoma uterus pCMV-SPORT6 L2265 NIH_MGC_39 adenocarcinoma pancreas pOTB7 L2269 NCI_CGAP_Thy11 follicular carcinoma thyroid pAMP10 L2270 Lupski_dorsal_root_ganglion dorsal root ganglia pCMV-SPORT6 (Life Technologies) L2285 BT0723 breast puc 18 L2289 BT0757 breast puc 18 L2293 BT0762 breast puc 18 L2300 BT0789 breast puc 18 L2323 CT0406 colon puc 18 L2336 CT0428 colon puc 18 L2352 UT0001 uterus_tumor puc 18 L2357 UT0021 uterus_tumor puc 18 L2359 UT0023 uterus_tumor puc 18 L2368 UT0041 uterus_tumor puc 18 L2380 NN0068 nervous_normal puc 18 L2381 NN0070 nervous_normal puc 18 L2402 NN0118 nervous_normal puc 18 L2412 NN0136 nervous_normal puc 18 L2439 NN1022 nervous_normal puc 18 L2482 HT0497 head_neck puc 18 L2486 HT0527 head_neck puc 18 L2487 HT0542 head_neck puc 18 L2491 HT0559 head_neck puc 18 L2497 HT0618 head_neck puc 18 L2499 HT0622 head_neck puc 18 L2504 HT0636 head_neck puc 18 L2519 HT0698 head_neck puc 18 L2528 HT0713 head_neck puc 18 L2543 HT0734 head_neck puc 18 L2551 HT0744 head_neck puc 18 L2630 HT0865 head_neck puc 18 L2635 HT0875 head_neck puc 18 L2647 HT0894 head_neck puc 18 L2652 NIH_MGC_57 glioblastoma brain pDNR-LIB (Clontech) L2653 NIH_MGC_58 hypernephroma kidney pDNR-LIB (Clontech) L2654 NIH_MGC_9 adenocarcinoma ovary pOTB7 cell line L2655 NIH_MGC_55 from acute myelogenous bone marrow pDNR-LIB leukemia (Clontech) L2657 NIH_MGC_54 from chronic myelogenous bone marrow pDNR-LIB leukemia (Clontech) L2669 NT0022 nervous_tumor puc 18 L2670 NT0023 nervous_tumor puc 18 L2673 NT0028 nervous_tumor puc 18 L2675 NT0033 nervous_tumor puc 18 L2706 NT0102 nervous_tumor puc 18 L2744 FT0004 prostate_tumor puc 18 L2758 FT0027 prostate_tumor puc 18 L2759 FT0028 prostate_tumor puc 18 L2767 FT0044 prostate_tumor puc 18 L2771 FT0050 prostate_tumor puc 18 L2777 FT0056 prostate_tumor puc 18 L2791 FT0077 prostate_tumor puc 18 L2800 FT0097 prostate_tumor puc 18 L2842 UM0009 uterus puc 18 L2877 AN0027 amnion_normal puc 18 L2879 AN0032 amnion_normal puc 18 L2903 BN0039 breast_normal puc 18 L2904 BN0042 breast_normal puc 18 L2906 BN0047 breast_normal puc 18 L2909 BN0067 breast_normal puc 18 L2910 BN0070 breast_normal puc 18 L2915 BN0098 breast_normal puc 18 L2985 BN0257 breast_normal puc 18 L2991 BN0264 breast_normal puc 18 L2999 BN0273 breast_normal puc 18 L3001 BN0275 breast_normal puc 18 L3012 BN0296 breast_normal puc 18 L3019 BN0303 breast_normal puc 18 L3092 ET0023 lung_tumor puc 18 L3109 ET0046 lung_tumor puc 18 L3117 ET0068 lung_tumor puc 18 L3119 ET0072 lung_tumor puc 18 L3180 MT0101 marrow puc 18 L3181 MT0107 marrow puc 18 L3210 OT0067 ovary puc 18 L3215 OT0083 ovary puc 18 L3271 FN0094 prostate_normal puc 18 L3278 FN0104 prostate_normal puc 18 L3280 FN0106 prostate_normal puc 18 L3311 FN0180 prostate_normal puc 18 L3327 SN0024 stomach_normal puc 18 L3357 TN0034 testis_normal puc 18 L3372 TN0068 testis_normal puc 18 L3378 TN0080 testis_normal puc 18 L3385 Homo sapiens HeLa HeLa L3387 GKB hepatocellular carcinoma pBluescript sk(-) L3388 GKC hepatocellular carcinoma pBluescript sk(-) L3391 NIH_MGC_53 carcinoma, cell line bladder pDNR-LIB (Clontech) L3450 CT0508 colon puc 18 L3485 GN0070 placenta_normal puc 18 L3499 HT0617 head_neck puc 18 L3503 HT0870 head_neck puc 18 L3516 HT0913 head_neck puc 18 L3560 TN0023 testis_normal puc 18 L3563 TN0037 testis_normal puc 18 L3566 TN0046 testis_normal puc 18 L3576 TN0086 testis_normal puc 18 L3585 TN0119 testis_normal puc 18 L3592 TN0129 testis_normal puc 18 L3630 UT0071 uterus_tumor puc 18 L3632 UT0074 uterus_tumor puc 18 L3642 ADA Adrenal gland pBluescript sk(-) L3643 ADB Adrenal gland pBluescript sk(−) L3644 ADC Adrenal gland pBluescript sk(-) L3645 Cu adrenal cortico adenoma for pBluescript sk(-) Cushing''s syndrome L3646 DCA pTriplEx2 L3647 Human HO-1 melanoma cells L3649 DCB pTriplEx2 L3651 FHTA hypothalamus pTriplEx2 L3652 FHTB hypothalamus pTriplEx2 L3653 HTB Hypothalamus pBluescript sk(-) L3655 HTC Hypothalamus pBluescript sk(−) L3657 HTF Hypothalamus pBluescript sk(-) L3658 cdA pheochromocytoma pTriplEx2 L3659 CB cord blood pBluescript L3660 NP1 pituitary pBluescript sk(-) L3661 NPA pituitary pBluescript sk(-) L3663 NIH_MGC_60 adenocarcinoma prostate pDNR-LIB (Clontech) L3665 NIH_MGC_75 kidney pDNR-LIB (Clontech) L3709 CT0515 colon puc 18 L3726 GN0038 placenta_normal puc 18 L3729 GN0079 placenta_normal puc 18 L3750 HT0945 head_neck puc 18 L3783 TN0136 testis_normal puc 18 L3796 UT0042 uterus_tumor puc 18 L3807 UT0077 uterus_tumor puc 18 L3811 NPC pituitary pBluescript sk(−) L3812 NPD pituitary pBluescript sk(-) L3813 TP pituitary tumor pTriplEx2 L3814 BM Bone marrow pTriplEx2 L3815 MDS Bone marrow pTriplEx2 L3816 HEMBA1 whole embryo, mainly head pME18SFL3 L3817 HEMBB1 whole embryo, mainly body pME18SFL3 L3818 MAMMA1 mammary gland pME18SFL3 L3820 NIH_MGC_46 leiomyosarcoma cell line uterus pOTB7 L3821 NIH_MGC_48 primary B-cells from tonsils B-cells pOTB7 (cell line) L3822 NIH_MGC_59 mucoepidermoid carcinoma lung pDNR-LIB (Clontech) L3823 NT2RM1 NT2 pUC19FL3 L3824 NT2RM2 NT2 pME18SFL3 L3825 NT2RM4 NT2 pME18SFL3 L3826 NT2RP1 NT2 pUC19FL3 L3827 NT2RP2 NT2 pME18SFL3 L3828 NT2RP3 NT2 pME18SFL3 L3829 NT2RP4 NT2 pME18SFL3 L3831 OVARC1 ovary, tumor tissue pME18SFL3 L3832 PLACE1 placenta pME18SFL3 L3833 PLACE2 placenta pME18SFL3 L3834 PLACE3 placenta pME18SFL3 L3837 THYRO1 thyroid gland pME18SFL3 L3872 NCI_CGAP_Skn1 skin, normal, 4 pCMV-SPORT6 pooled sa L3904 NCI_CGAP_Brn64 glioblastoma with brain pCMV-SPORT6 EGFR amplification L3905 NCI_CGAP_Brn67 anaplastic brain pCMV-SPORT6 oligodendroglioma with 1p/19q loss L4497 NCI_CGAP_Br22 invasive ductal carcinoma, 3 breast pCMV-SPORT6 pooled samples L4500 NCI_CGAP_HN16 moderate to poorly mouth pAMP10 differentiated invasive carcino L4501 NCI_CGAP_Sub8 pT7T3D-Pac (Pharmacia) with a modified polylinker L4507 NCI_CGAP_Thy6 normal epithelium thyroid pAMP10 L4508 NCI_CGAP_Thy8 normal epithelium thyroid pAMP10 L4537 NCI_CGAP_Thy7 follicular adenoma (benign thyroid pAMP10 lesion) L4556 NCI_CGAP_HN13 squamous cell carcinoma tongue pCMV-SPORT6 L4557 NCI_CGAP_Adr1 neuroblastoma adrenal gland pCMV-SPORT6 L4558 NCI_CGAP_Pan3 pancreas pCMV-SPORT6 L4559 NCI_CGAP_Thy3 follicular carcinoma thyroid pCMV-SPORT6 L4560 NCI_CGAP_Ut7 tumor uterus pCMV-SPORT6 L4669 NCI_CGAP_Ov41 serous papillary tumor ovary pCMV-SPORT6 L4747 NCI_CGAP_Brn41 oligodendroglioma brain pT7T3D-Pac (Pharmacia) with a modified polylinker L4753 NCI_CGAP_HN15 leukoplakia of the buccal mouth pAMP10 mucosa L4775 NCI_CGAP_Thy12 papillary carcinoma thyroid pAMP10 L5286 NCI_CGAP_Thy10 medullary carcinoma thyroid pAMP10 L5564 NCI_CGAP_HN20 normal pAMP1 head/neck tissue L5565 NCI_CGAP_Brn66 glioblastoma with brain pCMV-SPORT6 probably TP53 mutation and witho L5566 NCI_CGAP_Brn70 anaplastic brain pCMV-SPORT6.ccdb oligodendroglioma L5568 NCI_CGAP_HN21 nasopharyngeal head/neck pAMP1 carcinoma L5569 NCI_CGAP_HN17 normal epithelium nasopharynx pAMP10 L5572 NCI_CGAP_Co27 adenocarcinoma (mucinous colon pAMP1 component) L5574 NCI_CGAP_HN19 normal epithelium nasopharynx pAMP10 L5575 NCI_CGAP_Brn65 glioblastoma without brain pCMV-SPORT6 EGFR amplification L5622 NCI_CGAP_Skn3 skin pCMV-SPORT6 L5623 NCI_CGAP_Skn4 squamous cell carcinoma skin pCMV-SPORT6 Description of Table 5

Table 5 provides a key to the OMIM reference identification numbers disclosed in Table 1B.1, column 9. OMIM reference identification numbers (Column 1) were derived from Online Mendelian Inheritance in Man (Online Mendelian Inheritance in Man, OMIM. McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University (Baltimore, Md.) and National Center for Biotechnology Information, National Library of Medicine, (Bethesda, Md.) 2000. World Wide Web URL: http://www.ncbi.nlm.nih.gov/omim/). Column 2 provides diseases associated with the cytologic band disclosed in Table 1B.1, column 8, as determined using the Morbid Map database. TABLE 5 OMIM Reference Description 100678 ACAT2 deficiency 100690 Myasthenic syndrome, slow-channel congenital, 601462 100710 Myasthenic syndrome, slow-channel congenital, 601462 100730 Myasthenia gravis, neonatal transient 101000 Meningioma, NF2-related, sporadic Schwannoma, sporadic 101000 Neurofibromatosis, type 2 101000 Neurolemmomatosis 101000 Malignant mesothelioma, sporadic 102200 Somatotrophinoma 102540 Cardiomyopathy, idiopathic dilated 102578 Leukemia, acute promyelocytic, PML/RARA type 102700 Severe combined immunodeficiency due to ADA deficiency 102700 Hemolytic anemia due to ADA excess 102770 Myoadenylate deaminase deficiency 102772 [AMP deaminase deficiency, erythrocytic] 103000 Hemolytic anemia due to adenylate kinase deficiency 103050 Autism, succinylpurinemic 103050 Adenylosuccinase deficiency 103581 Albright hereditary osteodystrophy-2 103600 [Dysalbuminemic hyperthyroxinemia] 103600 [Dysalbuminemic hyperzincemia], 194470 103600 Analbuminemia 103720 Alcoholism, susceptibility to 103850 Aldolase A deficiency 103950 Emphysema due to alpha-2-macroglobulin deficiency 104150 [AFP deficiency, congenital] 104150 [Hereditary persistence of alpha-fetoprotein] 104311 Alzheimer disease-3 104500 Amelogenesis imperfecta-2, hypoplastic local type 104770 Amyloidosis, secondary, susceptibility to 105580 Anal canal carcinoma 105600 Dyserythropoietic anemia, congenital, type III 106100 Angioedema, hereditary 106150 Hypertension, essential, susceptibility to 106150 Preeclampsia, susceptibility to 106165 Hypertension, essential, 145500 106180 Myocardial infarction, susceptibility to 106210 Peters anomaly 106210 Cataract, congenital, with late-onset corneal dystrophy 106210 Foveal hypoplasia, isolated, 136520 106210 Aniridia 106300 Ankylosing spondylitis 107250 Anterior segment mesenchymal dysgenesis 107271 CD59 deficiency 107300 Antithrombin III deficiency 107470 Atypical mycobacterial infection, familial disseminated, 209950 107470 BCG infection, generalized familial 107470 Tuberculosis, susceptibility to 107670 Apolipoprotein A-II deficiency 107680 ApoA-I and apoC-III deficiency, combined 107680 Corneal clouding, autosomal recessive 107680 Amyloidosis, 3 or more types 107680 Hypertriglyceridemia, one form 107680 Hypoalphalipoproteinemia 107720 Hypertriglyceridemia 107741 Hyperlipoproteinemia, type III 107777 Diabetes insipidus, nephrogenic, autosomal recessive, 222000 107970 Arrhythmogenic right ventricular dysplasia-1 108120 Distal arthrogryposis-1 108725 Atherosclerosis, susceptibility to 108730 Brody myopathy, 601003 108800 Atrial septal defect, secundum type 108962 Hypertension, salt-resistant 108985 Atrophia areata 109150 Machado-Joseph disease 109270 Renal tubular acidosis, distal, 179800 109270 Spherocytosis, hereditary 109270 [Acanthocytosis, one form] 109270 [Elliptocytosis, Malaysian-Melanesian type] 109270 Hemolytic anemia due to band 3 defect 109565 Lymphoma, B-cell 109565 Lymphoma, diffuse large cell 109690 Asthma, nocturnal, susceptibility to 109690 Obesity, susceptibility to 109700 Hemodialysis-related amyloidosis 110100 Blepharophimosis, epicanthus inversus, and ptosis, type 1 110700 Vivax malaria, susceptibility to 112250 Bone dysplasia with medullary fibrosarcoma 112262 Fibrodysplasia ossificans progressiva, 135100 112410 Hypertension with brachydactyly 113721 Breast cancer 113811 Epidermolysis bullosa, generalized atrophic benign, 226650 113900 Heart block, progressive familial, type I 114130 Osteoporosis 114208 Malignant hyperthermia susceptibility 5, 601887 114208 Hypokalemic periodic paralysis, 170400 114240 Muscular dystrophy, limb-girdle, type 2A, 253600 114290 Campomelic dysplasia with autosomal sex reversal 114350 Leukemia, acute myeloid 114400 Lynch cancer family syndrome II 114550 Hepatocellular carcinoma 114835 Monocyte carboxyesterase deficiency 115200 Cardiomyopathy, dilated, 1A 115500 Acatalasemia 115650 Cataract, anterior polar-1 115660 Cataract, cerulean, type 1 115665 Cataract, congenital, Volkmann type 116800 Cataract, Marner type 116806 Colorectal cancer 116860 Cavernous angiomatous malformations 117700 [Hypoceruloplasminemia, hereditary] 117700 Hemosiderosis, systemic, due to aceruloplasminemia 118210 Charcot-Marie-Tooth neuropathy-2A 118425 Myotonia congenita, dominant, 160800 118425 Myotonia congenita, recessive, 255700 118425 Myotonia levior, recessive 118485 Polycystic ovary syndrome with hyperandrogenemia 118504 Epilepsy, benign neonatal, type 1, 121200 118504 Epilepsy, nocturnal frontal lobe, 600513 118511 Schizophrenia, neurophysiologic defect in 118800 Choreoathetosis, familial paroxysmal 119300 van der Woude syndrome 120070 Alport syndrome, autosomal recessive, 203780 120110 Metaphyseal chondrodysplasia, Schmid type 120120 Epidermolysis bullosa dystrophica, dominant, 131750 120120 Epidermolysis bullosa dystrophica, recessive, 226600 120120 Epidermolysis bullosa, pretibial, 131850 120131 Alport syndrome, autosomal recessive, 203780 120131 Hematuria, familial benign 120140 Osteoarthrosis, precocious 120140 SED congenita 120140 SMED Strudwick type 120140 Stickler syndrome, type I 120140 Wagner syndrome, type II 120140 Achondrogenesis-hypochondrogenesis, type II 120140 Kniest dysplasia 120150 Osteogenesis imperfecta, 4 clinical forms, 166200, 166210, 259420, 166220 120150 Osteoporosis, idiopathic, 166710 120150 Ehlers-Danlos syndrome, type VIIA1, 130060 120160 Osteogenesis imperfecta, 4 clinical forms, 166200, 166210, 259420, 166220 120160 Osteoporosis, idiopathic, 166710 120160 Ehlers-Danlos syndrome, type VIIA2, 130060 120160 Marfan syndrome, atypical 120180 Ehlers-Danlos syndrome, type III 120180 Ehlers-Danlos syndrome, type IV, 130050 120180 Fibromuscular dysplasia of arteries, 135580 120180 Aneurysm, familial, 100070 120190 Ehlers-Danlos syndrome, type I, 130000 120215 Ehlers-Danlos syndrome, type I, 130000 120215 Ehlers-Danlos syndrome, type II, 130010 120220 Bethlem myopathy, 158810 120240 Bethlem myopathy, 158810 120260 Epiphyseal dysplasia, multiple, type 2, 600204 120280 Stickler syndrome, type III 120280 Marshall syndrome, 154780 120290 OSMED syndrome, 215150 120290 Stickler syndrome, type II, 184840 120435 Muir-Torre syndrome, 158320 120435 Colorectal cancer, hereditary, nonpolyposis, type 1 Ovarian cancer 120436 Muir-Torre family cancer syndrome, 158320 120436 Turcot syndrome with glioblastoma, 276300 120436 Colorectal cancer, hereditary nonpolyposis, type 2 120550 C1q deficiency, type A 120570 C1q deficiency, type B 120575 C1q deficiency, type C 120580 C1r/C1s deficiency, combined 120620 SLE susceptibility 120620 CR1 deficiency 120700 C3 deficiency 120810 C4 deficiency 120820 C4 deficiency 120900 C5 deficiency 120920 Measles, susceptibility to 120940 C9 deficiency 120950 C8 deficiency, type I 120960 C8 deficiency, type II 121014 Heterotaxia, visceroatrial, autosomal recessive 121050 Contractural arachnodactyly, congenital 121360 Myeloid leukemia, acute, M4Eo subtype 121800 Corneal dystrophy, crystalline, Schnyder 122720 Nicotine addiction, protection from 122720 Coumarin resistance, 122700 123000 Craniometaphyseal dysplasia 123100 Craniosynostosis, type 1 123101 Craniosynostosis, type 2 123270 [Creatine kinase, brain type, ectopic expression of] 123580 Cataract, congenital, autosomal dominant 123620 Cataract, cerulean, type 2, 601547 123660 Cataract, Coppock-like 123829 Melanoma 123940 White sponge nevus, 193900 124020 Mephenytoin poor metabolizer 124030 Parkinsonism, susceptibility to 124030 Debrisoquine sensitivity 124200 Darier disease (keratosis follicularis) 125264 Leukemia, acute nonlymphocytic 125270 Porphyria, acute hepatic 125270 Lead poisoning, susceptibility to 125370 Dentatorubro-pallidoluysian atrophy 125490 Dentinogenesis imperfecta-1 125660 Myopathy, desminopathic 125660 Cardiomyopathy 125852 Insulin-dependent diabetes mellitus-2 126060 Anemia, megaloblastic, due to DHFR deficiency 126337 Myxoid liposarcoma 126340 Xeroderma pigmentosum, group D, 278730 126391 DNA ligase I deficiency 126451 Schizophrenia, susceptibility to 126452 Autonomic nervous system dysfunction 126452 [Novelty seeking personality] 126600 Drusen, radial, autosomal dominant 126650 Chloride diarrhea, congenital, Finnish type, 214700 126650 Colon cancer 128100 Dystonia-1, torsion 129010 Neuropathy, congenital hypomyelinating, 1 129490 Ectodermal dysplasia-3, anhidrotic 129900 EEC syndrome-1 130410 Glutaricaciduria, type IIB 130500 Elliptocytosis-1 130650 Beckwith-Wiedemann syndrome 131100 Multiple endocrine neoplasia I 131100 Prolactinoma, hyperparathyroidism, carcinoid syndrome 131100 Carcinoid tumor of lung 131195 Hereditary hemorrhagic telangiectasia-1, 187300 131210 Atherosclerosis, susceptibility to 131242 Shah-Waardenburg syndrome, 277580 131400 Eosinophilia, familial 131440 Eosinophilic myeloproliferative disorder 132700 Cylindromatosis 132810 Diphenylhydantoin toxicity 132810 Fetal hydantoin syndrome 133170 Erythremia 133171 [Erythrocytosis, familial], 133100 133200 Erythrokeratodermia variabilis 133430 Breast cancer 133430 Estrogen resistance 133450 Neuroepithelioma 133450 Ewing sarcoma 133510 Trichothiodystrophy 133510 Xeroderma pigmentosum, group B 133550 Dicarboxylicaminoaciduria, 222730 133700 Chondrosarcoma, 215300 133700 Exostoses, multiple, type 1 133701 Exostoses, multiple, type 2 133780 Vitreoretinopathy, exudative, familial 134370 Membroproliferative glomerulonephritis 134370 Factor H deficiency 134370 Hemolytic-uremic syndrome, 235400 134570 Factor XIIIA deficiency 134580 Factor XIIIB deficiency 134637 Autoimmune lymphoproliferative syndrome 134790 Hyperferritinemia-cataract syndrome, 600886 134820 Dysfibrinogenemia, alpha type, causing bleeding diathesis 134820 Dysfibrinogenemia, alpha type, causing recurrent thrombosis 134820 Amyloidosis, hereditary renal, 105200 134830 Dysfibrinogenemia, beta type 134850 Dysfibrinogenemia, gamma type 134850 Hypofibrinogenemia, gamma type 134934 Thanatophoric dysplasia, types I and II, 187600 134934 Achondroplasia, 100800 134934 Craniosynostosis, nonsyndromic 134934 Crouzon syndrome with acanthosis nigricans 134934 Hypochondroplasia, 146000 135300 Fibromatosis, gingival 135600 Ehlers-Danlos syndrome, type X 135700 Fibrosis of extraocular muscles, congenital, 1 135940 Ichthyosis vulgaris, 146700 136132 [Fish-odor syndrome], 602079 136350 Pfeiffer syndrome, 101600 136435 Ovarian dysgenesis, hypergonadotropic, with normal karyotype, 233300 136530 Male infertility, familial 136836 Fucosyltransferase-6 deficiency 137350 Amyloidosis, Finnish type, 105120 138030 [Hyperproglucagonemia] 138033 Diabetes mellitus, type II 138040 Cortisol resistance 138079 Hyperinsulinism, familial, 602485 138079 MODY, type 2, 125851 138130 Hyperinsulinism-hyperammonemia syndrome 138140 Glucose transport defect, blood-brain barrier 138160 Diabetes mellitus, noninsulin-dependent 138160 Fanconi-Bickel syndrome, 227810 138190 Diabetes mellitus, noninsulin-dependent 138250 P5CS deficiency 138300 Hemolytic anemia due to glutathione reductase deficiency 138320 Hemolytic anemia due to glutathione peroxidase deficiency 138491 Startle disease, autosomal recessive 138491 Startle disease/hyperekplexia, autosomal dominant, 149400 138491 Hyperekplexia and spastic paraparesis 138570 Non-insulin dependent diabetes mellitus, susceptibility to 138571 Glycogen synthase, liver, deficiency of, 240600 138700 [Apolipoprotein H deficiency] 138720 Bernard-Soulier syndrome, type B 138971 Kostmann neutropenia, 202700 138981 Pulmonary alveolar proteinosis, 265120 139130 Hypertension, essential, susceptibility to, 145500 139150 Basal cell carcinoma 139190 Gigantism due to GHRF hypersecretion 139190 Isolated growth hormone deficiency due to defect in GHRF 139191 Growth hormone deficient dwarfism 139250 Isolated growth hormone deficiency, Illig type with absent GH and Kowarski type with bioinactive GH 139320 Pituitary ACTH secreting adenoma 139320 Pseudohypoparathyroidism, type Ia, 103580 139320 Somatotrophinoma 139320 McCune-Albright polyostotic fibrous dysplasia, 174800 139330 Night blindness, congenital stationary 139350 Epidermolytic hyperkeratosis, 113800 139350 Keratoderma, palmoplantar, nonepidermolytic 139360 Pituitary ACTH-secreting adenoma 140100 [Anhaptoglobinemia] 140100 [Hypohaptogloginemia] 141750 Alpha-thalassemia/mental retardation syndrome, type 1 141800 Methemoglobinemias, alpha- 141800 Thalassemias, alpha- 141800 Erythremias, alpha- 141800 Heinz body anemias, alpha- 141850 Thalassemia, alpha- 141850 Erythrocytosis 141850 Heinz body anemia 141850 Hemoglobin H disease 141850 Hypochromic microcytic anemia 141900 Methemoglobinemias, beta- 141900 Sickle cell anemia 141900 Thalassemias, beta- 141900 Erythremias, beta- 141900 HPFH, deletion type 141900 Heinz body anemias, beta- 142000 Thalassemia due to Hb Lepore 142000 Thalassemia, delta- 142200 HPFH, nondeletion type A 142250 HPFH, nondeletion type G 142270 Hereditary persistence of fetal hemoglobin 142335 Hereditary persistence of fetal hemoglobin, heterocellular, Indian type 142470 [Hereditary persistence of fetal hemoglobin, heterocellular] 142640 Thrombophilia due to elevated HRG 142680 Periodic fever, familial 142857 Pemphigoid, susceptibility to 142858 Beryllium disease, chronic, susceptibility to 142946 Holoprosencephaly-4 142959 Hand-foot-uterus syndrome, 140000 142989 Synpolydactyly, type II, 186000 143100 Huntington disease 143200 Wagner syndrome 143200 Erosive vitreoretinopathy 143890 Hypercholesterolemia, familial 144200 Epidermolytic palmoplantar keratoderma 145001 Hyperparathyroidism-jaw tumor syndrome 145260 Pseudohypoaldosteronism, type II 145410 Opitz G syndrome, type II 145505 Hypertension, essential 145981 Hypocalciuric hypercalcemia, type II 146150 Hypomelanosis of Ito 146150 Hypomelanosis of Ito 146760 [IgG receptor I, phagocytic, familial deficiency of] 146790 Lupus nephritis, susceptibility to 147050 Atopy 147141 Leukemia, acute lymphoblastic 147200 [Kappa light chain deficiency] 147280 Hepatocellular carcinoma 147440 Growth retardation with deafness and mental retardation 147450 Amytrophic lateral sclerosis, due to SOD1 deficiency, 105400 147545 Diabetes mellitus, noninsulin-dependent 147570 Interferon, immune, deficiency 147660 Interferon, alpha, deficiency 147670 Rabson-Mendenhall syndrome 147670 Diabetes mellitus, insulin-resistant, with acanthosis nigricans 147670 Leprechaunism 147730 Interleukin-2 receptor, alpha chain, deficiency of 147781 Atopy, susceptibility to 147790 Leukemia, acute lymphocytic, with 4/11 translocation 147791 Jacobsen syndrome 148040 Epidermolysis bullosa simplex, Koebner, Dowling-Meara, and Weber- Cockayne types, 131900, 131760, 131800 148041 Pachyonychia congenita, Jadassohn-Lewandowsky type, 167200 148043 Meesmann corneal dystrophy, 122100 148065 White sponge nevus, 193900 148066 Epidermolysis bullosa simplex, Koebner, Dowling-Meara, and Weber- Cockayne types, 131900, 131760, 131800 148066 Epidermolysis bullosa simplex, recessive, 601001 148067 Nonepidermolytic palmoplantar keratoderma, 600962 148067 Pachyonychia congenita, Jadassohn-Lewandowsky type, 167200 148069 Pachyonychia congenita, Jackson-Lawler type, 167210 148070 Liver disease, susceptibility to, from hepatotoxins or viruses 148080 Epidermolytic hyperkeratosis, 113800 148370 Keratolytic winter erythema 148500 Tylosis with esophageal cancer 150000 Exertional myoglobinuria due to deficiency of LDH-A 150100 Lactate dehydrogenase-B deficiency 150200 [Placental lactogen deficiency] 150210 Lactoferrin-deficient neutrophils, 245480 150230 Langer-Giedion syndrome 150240 Cutis laxa, marfanoid neonatal type 150250 Larsen syndrome, autosomal dominant 150270 Laryngeal adductor paralysis 150292 Epidermolysis bullosa, Herlitz junctional type, 226700 150310 Epidermolysis bullosa, Herlitz junctional type, 226700 150310 Epidermolysis bullosa, generalized atrophic benign, 226650 151385 Leukemia, acute myeloid 151390 Leukemia, acute T-cell 151400 Leukemia/lymphoma, B-cell, 1 151410 Leukemia, chronic myeloid 151440 Leukemia, T-cell acute lymphoblastoid 151670 Hepatic lipase deficiency 152200 Coronary artery disease, susceptibility to 152427 Long QT syndrome-2 152445 Vohwinkel syndrome, 124500 152445 Erythrokeratoderma, progressive symmetric, 602036 152760 Hypogonadotropic hypogonadism due to GNRH deficiency, 227200 152790 Precocious puberty, male, 176410 152790 Leydig cell hypoplasia 153454 Ehlers-Danlos syndrome, type VI, 225400 153455 Cutis laxa, recessive, type I, 219100 153700 Macular dystrophy, vitelliform type 153880 Macular dystrophy, dominant cystoid 153900 Stargardt disease-2 154275 Malignant hyperthermia susceptibility 2 154276 Malignant hyperthermia susceptibility 3 154400 Acrofacial dysostosis, Nager type 154500 Treacher Collins mandibulofacial dysostosis 154550 Carbohydrate-deficient glycoprotein syndrome, type Ib, 602579 154705 Marfan syndrome, type II 155555 [Red hair/fair skin] 155555 UV-induced skin damage, vulnerability to 155600 Malignant melanoma, cutaneous 156225 Muscular dystrophy, congenital merosin-deficient 156232 Mesomelic dysplasia, Kantaputra type 156570 Methylcobalamin deficiency, cb1 G type 156600 Microcoria, congenital 156845 Tietz syndrome, 103500 156845 Waardenburg syndrome, type IIA, 193510 156845 Waardenburg syndrome/ocular albinism, digenic, 103470 156850 Cataract, congenital, with microphthalmia 157147 Abetalipoproteinemia, 200100 157170 Holoprosencephaly-2 157640 PEO with mitochondrial DNA deletions, type 1 157655 Lactic acidosis due to defect in iron-sulfur cluster of complex I 157900 Moebius syndrome 159000 Muscular dystrophy, limb-girdle, type 1A 159001 Muscular dystrophy, limb-girdle, type 1B 159440 Charcot-Marie-Tooth neuropathy-1B, 118200 159440 Dejerine-Sottas disease, myelin P-related, 145900 159440 Hypomyelination, congenital 159555 Leukemia, myeloid/lymphoid or mixed-lineage 160777 Griscelli disease, 214450 160781 Cardiomyopathy, hypertrophic, mid-left ventricular chamber type 160900 Myotonic dystrophy 161015 Mitochondrial complex I deficiency, 252010 162100 Neuralgic amyotrophy with predilection for brachial plexus 162200 Neurofibromatosis, type 1 162200 Watson syndrome, 193520 162400 Neuropathy, hereditary sensory and autonomic, type 1 163729 Hypertension, pregnancy-induced 163890 Parkinson disease, type 1, 601508 164009 Leukemia, acute promyelocytic, NUMA/RARA type 164040 Leukemia, acute promyelocytic, NPM/RARA type 164160 Obesity, severe, due to leptin deficiency 164200 Oculodentodigital dysplasia 164200 Syndactyly, type III, 186100 164500 Spinocerebellar ataxia-7 164731 Ovarian carcinoma, 167000 164759 Ovarian carcinoma 164790 Colorectal cancer 164860 Renal cell carcinoma, papillary, familial and sporadic 164920 Piebaldism 164920 Mast cell leukemia 164920 Mastocytosis with associated hematologic disorder 164953 Liposarcoma 165240 Pallister-Hall syndrome, 146510 165240 Postaxial polydactyly type A1, 174200 165240 Greig cephalopolysyndactyly syndrome, 175700 165320 Hepatocellular carcinoma 165500 Optic atrophy 1 166600 Osteopetrosis, AD, type II 167000 Ovarian cancer, serous 167250 Paget disease of bone 167409 Optic nerve coloboma with renal disease, 120330 167410 Rhabdomyosarcoma, alveolar, 268220 167415 Hypothyroidism, congenital, due to thyroid dysgenesis or hypoplasia 168000 Paraganglioma, familial nonchromaffin, 1 168360 Paraneoplastic sensory neuropathy 168450 Hypoparathyroidism, autosomal dominant 168450 Hypoparathyroidism, autosomal recessive 168461 Multiple myeloma, 254250 168461 Parathyroid adenomatosis 1 168461 Centrocytic lymphoma 168468 Metaphyseal chondrodysplasia, Murk Jansen type, 156400 168470 Humoral hypercalcemia of malignancy 168500 Parietal foramina 169600 Hailey-Hailey disease 170261 Bare lymphocyte syndrome, type I, due to TAP2 deficiency 170500 Myotonia congenita, atypical acetazolamide-responsive 170500 Paramyotonia congenita, 168300 170500 Hyperkalemic periodic paralysis 170650 Periodontitis, juvenile 170995 Zellweger syndrome-2 171050 Colchicine resistance 171060 Cholestasis, progressive familial intrahepatic, type III, 602347 171190 Hypertension, essential, 145500 171650 Lysosomal acid phosphatase deficiency 171760 Hypophosphatasia, adult, 146300 171760 Hypophosphatasia, infantile, 241500 171860 Hemolytic anemia due to phosphofructokinase deficiency 172400 Hemolytic anemia due to glucosephosphate isomerase deficiency 172400 Hydrops fetalis, one form 172411 Colorectal cancer, resistance to 172430 Enolase deficiency 172471 Glycogenosis, hepatic, autosomal 172490 Phosphorylase kinase deficiency of liver and muscle, 261750 173350 Plasminogen Tochigi disease 173350 Plasminogen deficiency, types I and II 173350 Thrombophilia, dysplasminogenemic 173360 Thrombophilia due to excessive plasminogen activator inhibitor 173360 Hemorrhagic diathesis due to PAI1 deficiency 173370 Plasminogen activator deficiency 173470 Glanzmann thrombasthenia, type B 173610 Platelet alpha/delta storage pool deficiency 173850 Polio, susceptibility to 173870 Xeroderma pigmentosum 173870 Fanconi anemia 173910 Polycystic kidney disease, adult, type II 174000 Medullary cystic kidney disease, AD 174810 Osteolysis, familial expansile 174900 Polyposis, juvenile intestinal 175100 Turcot syndrome, 276300 175100 Adenomatous polyposis coli 175100 Adenomatous polyposis coli, attenuated 175100 Colorectal cancer 175100 Desmoid disease, hereditary, 135290 175100 Gardner syndrome 176000 Porphyria, acute intermittent 176100 Porphyria cutanea tarda 176100 Porphyria, hepatoerythropoietic 176260 Episodic ataxia/myokymia syndrome, 160120 176261 Jervell and Lange-Nielsen syndrome, 220400 176270 Prader-Willi syndrome 176300 [Dystransthyretinemic hyperthyroxinemia] 176300 Carpal tunnel syndrome, familial 176300 Amyloid neuropathy, familial, several allelic types 176300 Amyloidosis, senile systemic 176450 Sacral agenesis-1 176730 Diabetes mellitus, rare form 176730 Hyperproinsulinemia, familial 176730 MODY, one form 176801 Metachromatic leukodystrophy due to deficiency of SAP-1 176801 Gaucher disease, variant form 176830 Obesity, adrenal insufficiency, and red hair 176830 ACTH deficiency 176860 Purpura fulminans, neonatal 176860 Thrombophilia due to protein C deficiency 176880 Protein S deficiency 176930 Dysprothrombinemia 176930 Hypoprothrombinemia 176943 Apert syndrome, 101200 176943 Pfeiffer syndrome, 101600 176943 Beare-Stevenson cutis gyrata syndrome, 123790 176943 Crouzon craniofacial dysostosis, 123500 176943 Jackson-Weiss syndrome, 123150 176947 Selective T-cell defect 176960 Pituitary tumor, invasive 177070 Spherocytosis, hereditary, Japanese type 177070 Hermansky-Pudlak syndrome, 203300 177400 Apnea, postanesthetic 177900 Psoriasis susceptibility-1 178300 Ptosis, hereditary congenital, 1 178600 Pulmonary hypertension, familial primary 178640 Pulmonary alveolar proteinosis, congenital, 265120 179095 Male infertility 179450 Ragweed sensitivity 179605 Retinitis pigmentosa, digenic 179605 Retinitis pigmentosa-7, peripherin-related 179605 Retinitis punctata albescens 179605 Butterfly dystrophy, retinal 179605 Macular dystrophy 179615 Reticulosis, familial histiocytic, 267700 179615 Severe combined immunodeficiency, B cell-negative, 601457 179616 Severe combined immunodeficiency, B cell-negative, 601457 179755 Renal cell carcinoma, papillary, 1 179820 [Hyperproreninemia] 180020 Retinal cone dystrophy-1 180069 Retinal dystrophy, autosomal recessive, childhood-onset 180069 Retinitis pigmentosa-20 180069 Leber congenital amaurosis-2, 204100 180071 Retinitis pigmentosa, autosomal recessive 180072 Night blindness, congenital stationary, type 3, 163500 180072 Retinitis pigmentosa, autosomal recessive 180100 Retinitis pigmentosa-1 180104 Retinitis pigmentosa-9 180105 Retinitis pigmentosa-10 180200 Osteosarcoma, 259500 180200 Pinealoma with bilateral retinoblastoma 180200 Retinoblastoma 180200 Bladder cancer, 109800 180240 Leukemia, acute promyelocytic 180250 Retinol binding protein, deficiency of 180297 Anemia, hemolytic, Rh-null, suppressor type, 268150 180380 Night blindness, congenital stationery, rhodopsin-related 180380 Retinitis pigmentosa, autosomal recessive 180380 Retinitis pigmentosa-4, autosomal dominant 180381 Oguchi disease-2, 258100 180385 Leukemia, acute T-cell 180721 Retinitis pigmentosa, digenic 180840 Susceptibility to IDDM 180860 Russell-Silver syndrome 180901 Malignant hyperthermia susceptibility 1, 145600 180901 Central core disease, 117000 181030 Salivary gland pleomorphic adenoma 181031 Oguchi disease-1, 258100 181405 Scapuloperoneal spinal muscular atrophy, New England type 181430 Scapuloperoneal syndrome, myopathic type 181460 Schistosoma mansoni, susceptibility/resistance to 181510 Schizophrenia 181600 Sclerotylosis 182138 Anxiety-related personality traits 182279 Prader-Willi syndrome 182280 Small-cell cancer of lung 182380 Glucose/galactose malabsorption 182381 Renal glucosuria, 253100 182452 Lung cancer, small cell 182500 Cataract, congenital 182600 Spastic paraplegia-3A 182601 Spastic paraplegia-4 182860 Pyropoikilocytosis 182860 Spherocytosis, recessive 182860 Elliptocytosis-2 182870 Spherocytosis-1 182870 Elliptocytosis-3 182870 Anemia, neonatal hemolytic, fatal and near-fatal 182900 Spherocytosis-2 183600 Split hand/foot malformation, type 1 185000 Stomatocytosis I 185430 Atherosclerosis, susceptibility to 185470 Myopathy due to succinate dehydrogenase deficiency 185800 Symphalangism, proximal 186580 Arthrocutaneouveal granulomatosis 186740 Immunodeficiency due to defect in CD3-gamma 186770 Leukemia, T-cell acute lymphocytic 186780 CD3, zeta chain, deficiency 186830 Immunodeficiency, T-cell receptor/CD3 complex 186860 Leukemia/lymphoma, T-cell 186880 Leukemia/lymphoma, T-cell 186921 Leukemia, T-cell acute lymphoblastic 187040 Leukemia-1, T-cell acute lymphoblastic 187680 6-mercaptopurine sensitivity 188025 Thrombocytopenia, Paris-Trousseau type 188070 Bleeding disorder due to defective thromboxane A2 receptor 188450 Goiter, adolescent multinodular 188450 Goiter, nonendemic, simple 188450 Hypothyroidism, hereditary congenital 188540 Hypothyroidism, nongoitrous 188826 Sorsby fundus dystrophy, 136900 189800 Preeclampsia/eclampsia 189980 Leukemia, chronic myeloid 190000 Atransferrinemia 190020 Bladder cancer, 109800 190040 Meningioma, SIS-related 190040 Dermatofibrosarcoma protuberans 190040 Giant-cell fibroblastoma 190100 Geniospasm 190160 Thyroid hormone resistance, 274300, 188570 190182 Colon cancer 190182 Colorectal cancer, familial nonpolyposis, type 6 190195 Ichthyosiform erythroderma, congenital, 242100 190195 Ichthyosis, lamellar, autosomal recessive, 242300 190198 Leukemia, T-cell acute lymphoblastic 190450 Hemolytic anemia due to triosephosphate isomerase deficiency 190605 Triphalangeal thumb-polysyndactyly syndrome 190685 Down syndrome 190900 Colorblindness, tritan 191010 Cardiomyopathy, familial hypertrophic, 3, 115196 191030 Nemaline myopathy-1, 161800 191044 Cardiomyopathy, familial hypertrophic 191045 Cardiomyopathy, familial hypertrophic, 2, 115195 191092 Tuberous sclerosis-2 191100 Tuberous sclerosis-1 191170 Colorectal cancer, 114500 191170 Li-Fraumeni syndrome 191181 Cervical carcinoma 191290 Segawa syndrome, recessive 191315 Insensitivity to pain, congenital, with anhidrosis, 256800 191540 [Urate oxidase deficiency] 192090 Ovarian carcinoma 192090 Breast cancer, lobular 192090 Endometrial carcinoma 192090 Gastric cancer, familial, 137215 192340 Diabetes insipidus, neurohypophyseal, 125700 192500 Jervell and Lange-Nielsen syndrome, 220400 192500 Long QT syndrome-1 192974 Neonatal alloimmune thrombocytopenia 192974 Glycoprotein Ia deficiency 193100 Hypophosphatemic rickets, autosomal dominant 193235 Vitreoretinopathy, neovascular inflammatory 193300 Renal cell carcinoma 193300 von Hippel-Lindau syndrome 193400 von Willebrand disease 193500 Rhabdomyosarcoma, alveolar, 268220 193500 Waardenburg syndrome, type I 193500 Waardenburg syndrome, type III, 148820 193500 Craniofacial-deafness-hand syndrome, 122880 194070 Wilms tumor, type 1 194070 Denys-Drash syndrome 194070 Frasier syndrome, 136680 194071 Wilms tumor, type 2 194071 Adrenocortical carcinoma, hereditary, 202300 194190 Wolf-Hirschhorn syndrome 200150 Choreoacanthocytosis 200990 Acrocallosal syndrome 201450 Acyl-CoA dehydrogenase, medium chain, deficiency of 201460 Acyl-CoA dehydrogenase, long chain, deficiency of 201470 Acyl-CoA dehydrogenase, short-chain, deficiency of 201910 Adrenal hyperplasia, congenital, due to 21-hydroxylase deficiency 202110 Adrenal hyperplasia, congenital, due to 17-alpha-hydroxylase deficiency 203100 Waardenburg syndrome/ocular albinism, digenic, 103470 203100 Albinism, oculocutaneous, type IA 203200 Albinism, ocular, autosomal recessive 203200 Albinism, oculocutaneous, type II 203300 Hermansky-Pudlak syndrome 203310 Ocular albinism, autosomal recessive 203500 Alkaptonuria 203740 Alpha-ketoglutarate dehydrogenase deficiency 203750 3-ketothiolase deficiency 203800 Alstrom syndrome 204500 Ceroid-lipofuscinosis, neuronal 2, classic late infantile 205100 Amyotrophic lateral sclerosis, juvenile 207750 Hyperlipoproteinemia, type Ib 207800 Argininemia 208100 Arthrogryposis multiplex congenita, neurogenic 208250 Jacobs syndrome 208400 Aspartylglucosaminuria 209901 Bardet-Biedl syndrome 1 211420 Breast cancer, ductal 212138 Carnitine-acylcarnitine translocase deficiency 214300 Klippel-Feil syndrome 214400 Charcot-Marie-Tooth neuropathy-4A 214500 Chediak-Higashi syndrome 215700 Citrullinemia 216550 Cohen syndrome 216900 Achromatopsia 216950 C1r/C1s deficiency, combined 217000 C2 deficiency 217050 C6 deficiency 217050 Combined C6/C7 deficiency 217070 C7 deficiency 217800 Macular corneal dystrophy 218000 Andermann syndrome 218030 Apparent mineralocorticoid excess, hypertension due to 219800 Cystinosis, nephropathic 221770 Polycystic lipomembranous osteodysplasia with sclerosing leukencephalopathy 221820 Gliosis, familial progressive subcortical 222100 Diabetes mellitus, insulin-dependent-1 222600 Atelosteogenesis II, 256050 222600 Achondrogenesis Ib, 600972 222600 Diastrophic dysplasia 222700 Lysinuric protein intolerance 222800 Hemolytic anemia due to bisphosphoglycerate mutase deficiency 222900 Sucrose intolerance 223000 Lactase deficiency, adult, 223100 223000 Lactase deficiency, congenital 223360 Dopamine-beta-hydroxylase deficiency 223900 Dysautonomia, familial 224100 Congenital dyserythropoietic anemia II 224120 Dyserythropoietic anemia, contenital, type I 225500 Ellis-van Creveld syndrome 227220 [Eye color, brown] 227500 Factor VII deficiency 227600 Factor X deficiency 227646 Fanconi anemia, type D 227650 Fanconi anemia, type A 228960 [Kininogen deficiency] 229300 Friedreich ataxia 229300 Friedreich ataxia with retained reflexes 229600 Fructose intolerance 229700 Fructose-bisphosphatase deficiency 229800 [Fructosuria] 230000 Fucosidosis 230200 Galactokinase deficiency with cataracts 230350 Galactose epimerase deficiency 230400 Galactosemia 230450 Hemolytic anemia due to gamma-glutamylcysteine synthetase deficiency 230800 Gaucher disease 230800 Gaucher disease with cardiovascular calcification 231550 Achalasia-addisonianism-alacrimia syndrome 231670 Glutaricaciduria, type I 231680 Glutaricaciduria, type IIA 231950 Glutathioninuria 232000 Propionicacidemia, type I or pccA type 232050 Propionicacidemia, type II or pccB type 232300 Glycogen storage disease II 232400 Glycogen storage disease IIIa 232400 Glycogen storage disease IIIb 232500 Glycogen storage disease IV 232600 McArdle disease 232700 Glycogen storage disease VI 232800 Glycogen storage disease VII 233100 [Renal glucosuria] 233700 Chronic granulomatous disease due to deficiency of NCF-1 233710 Chronic granulomatous disease due to deficiency of NCF-2 234000 Factor XII deficiency 234200 Neurodegeneration with brain iron accumulation 235200 Hemochromatosis 235800 [Histidinemia] 236100 Holoprosencephaly-1 236200 Homocystinuria, B6-responsive and nonresponsive types 236730 Urofacial syndrome 237300 Carbamoylphosphate synthetase I deficiency 238300 Hyperglycinemia, nonketotic, type I 238310 Hyperglycinemia, nonketotic, type II 238600 Chylomicronemia syndrome, familial 238600 Combined hyperlipemia, familial 238600 Hyperlipoproteinemia I 238600 Lipoprotein lipase deficiency 238970 HHH syndrome 239100 Van Buchem disease 239500 Hyperprolinemia, type I 240300 Autoimmune polyglandular disease, type I 240400 Scurvy 243500 Isovalericacidemia 245000 Papillon-Lefevre syndrome 245050 Ketoacidosis due to SCOT deficiency 245200 Krabbe disease 245349 Lacticacidemia due to PDX1 deficiency 245900 Norum disease 245900 Fish-eye disease 246450 HMG-CoA lyase deficiency 246530 Leukotriene C4 synthase deficiency 246600 Pancreatic lipase deficiency 246900 Lipoamide dehydrogenase deficiency 247200 Miller-Dieker lissencephaly syndrome 247640 Leukemia, acute lymphoblastic 248510 Mannosidosis, beta- 248600 Maple syrup urine disease, type Ia 248610 Maple syrup urine disease, type II 248611 Maple syrup urine disease, type Ib 249000 Meckel syndrome 249270 Thiamine-responsive megaloblastic anemia 250100 Metachromatic leukodystrophy 250250 Cartilage-hair hypoplasia 250790 Methemoglobinemia due to cytochrome b5 deficiency 250800 Methemoglobinemia, type I 250800 Methemoglobinemia, type II 251000 Methylmalonicaciduria, mutase deficiency type 251600 Microphthalmia, autosomal recessive 252500 Mucolipidosis II 252500 Mucolipidosis III 252800 Mucopolysaccharidosis Ih 252800 Mucopolysaccharidosis Ih/s 252800 Mucopolysaccharidosis Is 252900 Sanfilippo syndrome, type A 252940 Sanfilippo syndrome, type D 253000 Mucopolysaccharidosis IVA 253200 Maroteaux-Lamy syndrome, several forms 253250 Mulibrey nanism 253270 Multiple carboxylase deficiency, biotin-responsive 253601 Miyoshi myopathy, 254130 253601 Muscular dystrophy, limb-girdle, type 2B 253800 Walker-Warburg syndrome, 236670 253800 Fukuyama type congenital muscular dystrophy 254210 Myasthenia gravis, familial infantile 254770 Epilepsy, juvenile myoclonic 254780 Myoclonus epilepsy, Lafora type 255800 Schwartz-Jampel syndrome 256030 Nemaline myopathy-2 256100 Nephronophthisis, juvenile 256540 Galactosialidosis 256550 Sialidosis, type I 256550 Sialidosis, type II 256700 Neuroblastoma 256731 Ceroid-lipofuscinosis, neuronal-5, variant late infantile 256850 Giant axonal neuropathy-1 257200 Niemann-Pick disease, type A 257200 Niemann-Pick disease, type B 257220 Niemann-Pick disease, type C 257220 Niemann-Pick disease, type D, 257250 258501 3-methylglutaconicaciduria, type III 258870 Gyrate atrophy of choroid and retina with ornithinemia, B6 responsive or unresponsive 259700 Osteopetrosis, recessive 259730 Renal tubular acidosis-osteopetrosis syndrome 259770 Osteoporosis-pseudoglioma syndrome 259900 Hyperoxaluria, primary, type 1 261510 Pseudo-Zellweger syndrome 261515 Peroxisomal bifunctional enzyme deficiency 261600 Phenylketonuria 261600 [Hyperphenylalaninemia, mild] 261640 Phenylketonuria due to PTS deficiency 261670 Myopathy due to phosphoglycerate mutase deficiency 262000 Bjornstad syndrome 263200 Polycystic kidney disease, autosomal recessive 263700 Porphyria, congenital erythropoietic 264300 Pseudohermaphroditism, male, with gynecomastia 264470 Adrenoleukodystrophy, pseudoneonatal 264700 Pseudo-vitamin D dependency rickets 1 266100 Pyridoxine dependency with seizures 266150 Pyruvate carboxylase deficiency 266200 Anemia, hemolytic, due to PK deficiency 266300 [Hair color, red] 266600 Inflammatory bowel disease-1 267750 Knobloch syndrome 268900 [Sarcosinemia] 270100 Situs inversus viscerum 270800 Spastic paraplegia-5A 271245 Spinocerebellar ataxia-8, infantile, with sensory neuropathy 271900 Canavan disease 272750 GM2-gangliosidosis, AB variant 272800 Tay-Sachs disease 272800 [Hex A pseudodeficiency] 272800 GM2-gangliosidosis, juvenile, adult 273300 Male germ cell tumor 273800 Thrombocytopenia, neonatal alloimmune 273800 Glanzmann thrombasthenia, type A 274180 Thromboxane synthase deficiency 274270 Thymine-uraciluria 274270 Fluorouracil toxicity, sensitivity to 274600 Pendred syndrome 274600 Deafness, autosomal recessive 4 275350 Transcobalamin II deficiency 276000 Pancreatitis, hereditary, 167800 276000 Trypsinogen deficiency 276600 Tyrosinemia, type II 276700 Tyrosinemia, type I 276900 Usher syndrome, type 1A 276901 Usher syndrome, type 2 276902 Usher syndrome, type 3 276903 Usher syndrome, type 1B 276903 Deafness, autosomal dominant 11, neurosensory, 601317 276903 Deafness, autosomal recessive 2, neurosensory, 600060 276904 Usher syndrome, type 1C 277700 Werner syndrome 277730 Wernicke-Korsakoff syndrome, susceptibility to 278000 Wolman disease 278000 Cholesteryl ester storage disease 278250 Wrinkly skin syndrome 278300 Xanthinuria, type I 278700 Xeroderma pigmentosum, group A 278760 Xeroderma pigmentosum, group F 300008 Nephrolithiasis, type I, 310468 300008 Proteinuria, low molecular weight, with hypercalciuric nephrocalcinosis 300008 Dent disease, 300009 300008 Hypophosphatemia, type III 300011 Menkes disease, 309400 300011 Occipital horn syndrome, 304150 300011 Cutis laxa, neonatal 300031 Mental retardation, X-linked, FRAXF type 300032 Alpha-thalassemia/mental retardation syndrome, type 2, 301040 300032 Juberg-Marsidi syndrome, 309590 300037 Simpson dysmorphia syndrome, 312870 300039 Deafness, X-linked 3, conductive, with stapes fixation, 304400 300044 Wernicke-Korsakoff syndrome, susceptibility to 300046 Mental retardation, X-linked 23, nonspecific 300047 Mental retardation, X-linked 20 300048 Intestinal pseudoobstruction, neuronal, X-linked 300049 Nodular heterotopia, bilateral periventricular 300049 BPNH/MR syndrome 300055 Mental retardation with psychosis, pyramidal signs, and macroorchidism 300062 Mental retardation, X-linked 14 300071 Night blindness, congenital stationary, type 2 300076 Wood neuroimmunologic syndrome 300077 Mental retardation, X-linked 29 300088 Epilepsy, female restricted, with mental retardation 300100 Adrenoleukodystrophy 300100 Adrenomyeloneuropathy 300104 Mental retardation, X-linked nonspecific, 309541 300110 Night blindness, congenital stationary, X-linked incomplete, 300071 300123 Mental retardation with isolated growth hormone deficiency 300126 Dyskeratosis congenita-1, 305000 300136 Diabetes mellitus, insulin-dependent, X-linked, susceptibility to 300300 XLA and isolated growth hormone deficiency, 307200 300300 Agammaglobulinemia, type 1, X-linked 300600 Ocular albinism, Forsius-Eriksson type 301000 Thrombocytopenia, X-linked, 313900 301000 Wiskott-Aldrich syndrome 301200 Amelogenesis imperfecta 301201 Amelogenesis imperfecta-3, hypoplastic type 301300 Anemia, sideroblastic/hypochromic 301310 Anemia, sideroblastic, with spinocerebellar ataxia 301500 Fabry disease 301590 Anophthalmos-1 301830 Arthrogryposis, X-linked (spinal muscular atrophy, infantile, X-linked) 301835 Arts syndrome 301845 Bazex syndrome 301900 Borjeson-Forssman-Lehmann syndrome 302060 Noncompaction of left ventricular myocardium, isolated 302060 Barth syndrome 302060 Cardiomyopathy, X-linked dilated, 300069 302060 Endocardial fibroelastosis-2 302350 Nance-Horan syndrome 302960 Chondrodysplasia punctata, X-linked dominant 303400 Cleft palate, X-linked 303630 Alport syndrome, 301050 303630 Leiomyomatosis-nephropathy syndrome, 308940 303631 Leiomyomatosis, diffuse, with Alport syndrome 303700 Colorblindness, blue monochromatic 303800 Colorblindness, deutan 303900 Colorblindness, protan 304020 Cone dystrophy, progressive X-linked, 1 304040 Charcot-Marie-Tooth neuropathy, X-linked-1, dominant, 302800 304340 Mental retardation, X-linked, syndromic-5, with Dandy-Walker malformation, basal ganglia disease, and seizures 304500 Deafness, X-linked 2, perceptive congenital 304700 Mohr-Tranebjaerg syndrome 304700 Deafness, X-linked 1, progressive 304700 Jensen syndrome, 311150 304800 Diabetes insipidus, nephrogenic 305100 Anhidrotic ectodermal dysplasia 305400 Aarskog-Scott syndrome 305450 FG syndrome 305900 Favism 305900 G6PD deficiency 305900 Hemolytic anemia due to G6PD deficiency 306700 Hemophilia A 306900 Hemophilia B 306995 [Homosexuality, male] 307150 Hypertrichosis, congenital generalized 307700 Hypoparathyroidism, X-linked 308000 HPRT-related gout 308000 Lesch-Nyhan syndrome 308230 Immunodeficiency, X-linked, with hyper-IgM 308240 Lymphoproliferative syndrome, X-linked 308300 Incontinentia pigmenti, sporadic type 308310 Incontinentia pigmenti, familial 308380 Severe combined immunodeficiency, X-linked, 300400 308380 Combined immunodeficiency, X-linked, moderate, 312863 308840 Spastic paraplegia, 312900 308840 Hydrocephalus due to aqueductal stenosis, 307000 308840 MASA syndrome, 303350 309000 Lowe syndrome 309200 Manic-depressive illness, X-linked 309300 Megalocornea, X-linked 309470 Mental retardation, X-linked, syndromic-3, with spastic diplegia 309500 Renpenning syndrome-1 309545 Mental retardation, X-linked nonspecific, with aphasia 309548 Mental retardation, X-linked, FRAXE type 309555 Gustavson syndrome 309605 Mental retardation, X-linked, syndromic-4, with congenital contractures and low fingertip arches 309610 Mental retardation, X-linked, syndromic-2, with dysmorphism and cerebral atrophy 309620 Mental retardation-skeletal dysplasia 309850 Brunner syndrome 309900 Mucopolysaccharidosis II 310300 Emery-Dreifuss muscular dystrophy 310400 Myotubular myopathy, X-linked 310460 Myopia-1 310460 Bornholm eye disease 310490 Cowchock syndrome 310500 Night blindness, congenital stationary, type 1 311050 Optic atrophy, X-linked 311200 Oral-facial-digital syndrome 1 311300 Otopalatodigital syndrome, type I 311510 Waisman parkinsonism-mental retardation syndrome 311800 Myoglobinuria/hemolysis due to PGK deficiency 311800 Hemolytic anemia due to PGK deficiency 311850 Phosphoribosyl pyrophosphate synthetase-related gout 311870 Muscle glycogenosis 312000 Panhypopituitarism, X-linked 312040 N syndrome, 310465 312060 Properdin deficiency, X-linked 312080 Pelizaeus-Merzbacher disease 312080 Spastic paraplegia-2, 312920 312600 Retinitis pigmentosa-2 312760 Turner syndrome 313350 Split hand/foot malformation, type 2 313850 Thoracoabdominal syndrome 314200 [Euthyroidal hyper- and hypothyroxinemia] 314250 Dystonia-3, torsion, with parkinsonism, Filipino type 314300 Goeminne TKCR syndrome 314400 Cardiac valvular dysplasia-1 314580 Wieacker-Wolff syndrome 600020 Prostate cancer, 176807 600035 Schizencephaly 600040 Colorectal cancer 600044 Thrombocythemia, essential, 187950 600045 Xeroderma pigmentosum, group E, subtype 2 600048 Breast cancer-3 600059 Retinitis pigmentosa-13 600065 Leukocyte adhesion deficiency, 116920 600079 Colon cancer 600095 Split hand/foot malformation, type 3 600101 Deafness, autosomal dominant 2 600105 Retinitis pigmentosa-12, autosomal recessive 600119 Muscular dystrophy, Duchenne-like, type 2 600119 Adhalinopathy, primary 600138 Retinitis pigmentosa-11 600140 Rubenstein-Taybi syndrome, 180849 600143 Epilepsy, progressive, with mental retardation 600151 Bardet-Biedl syndrome 3 600160 Melanoma, 155601 600163 Long QT syndrome-3 600173 SCID, autosomal recessive, T-negative/B-positive type 600175 Spinal muscular atrophy, congenital nonprogressive, of lower limbs 600179 Leber congenital amaurosis, type I, 204000 600184 Carnitine acetyltransferase deficiency 600185 Pancreatic cancer 600185 Breast cancer 2, early onset 600194 Ichthyosis bullosa of Siemens, 146800 600202 Dyslexia, specific, 2 600211 Cleidocranial dysplasia, 119600 600221 Venous malformations, multiple cutaneous and mucosal, 600195 600223 Spinocerebellar ataxia-4 600225 Phenylketonuria, atypical, due to GCH1 deficiency, 233910 600225 Dystonia, DOPA-responsive, 128230 600228 Pseudohypoaldosteronism, type I, 264350 600231 Palmoplantar keratoderma, Bothnia type 600234 HMG-CoA synthease-2 deficiency 600243 Temperature-sensitive apoptosis 600258 Colorectal cancer, hereditary nonpolyposis, type 3 600259 Turcot syndrome with glioblastoma, 276300 600259 Colorectal cancer, hereditary nonpolyposis, type 4 600261 Ehlers-Danlos-like syndrome 600266 Resistance/susceptibility to TB, etc. 600273 Polycystic kidney disease, infantile severe, with tuberous sclerosis 600276 Cerebral arteriopathy with subcortical infarcts and leukoencephalopathy, 125310 600281 Non-insulin-dependent diabetes mellitus, 125853 600281 MODY, type 1, 125850 600309 Atrioventricular canal defect-1 600310 Pseudoachondroplasia, 177170 600310 Epiphyseal dysplasia, multiple 1, 132400 600319 Diabetes mellitus, insulin-dependent, 4 600320 Insulin-dependent diabetes mellitus-5 600321 Diabetes mellitus, insulin-dependent, 7 600332 Rippling muscle disease-1 600354 Spinal muscular atrophy-1, 253300 600354 Spinal muscular atrophy-2, 253550 600354 Spinal muscular atrophy-3, 253400 600364 Cone dystrophy-3, 602093 600374 Bardet-Biedl syndrome 4 600414 Adrenoleukodystrophy, neonatal, 202370 600415 Ataxia with isolated vitamin E deficiency, 277460 600429 [Ii blood group, 110800] 600509 Persistent hyperinsulinemic hypoglycemia of infancy, 256450 600510 Pigment dispersion syndrome 600511 Schizophrenia-3 600512 Epilepsy, partial 600525 Trichodontoosseous syndrome, 190320 600528 CPT deficiency, hepatic, type I, 255120 600536 Myopathy, congenital 600542 Chondrosarcoma, extraskeletal myxoid 600584 Atrial septal defect with atrioventricular conduction defects, 108900 600593 Craniosynostosis, Adelaide type 600617 Lipoid adrenal hyperplasia, 201710 600618 Leukemia, acute lymphoblastic 600623 Prostate cancer, 176807 600624 Cone-rod retinal dystrophy-1 600631 Enuresis, nocturnal, 1 600650 Myopathy due to CPT II deficiency, 255110 600650 CPT deficiency, hepatic, type II, 600649 600698 Salivary adenoma 600698 Uterine leiomyoma 600698 Lipoma 600698 Lipomatosis, mutiple, 151900 600700 Lipoma 600701 Lipoma 600722 Ceroid lipofuscinosis, neuronal, variant juvenile type, with granular osmiophilic deposits 600722 Ceroid lipofuscinosis, neuronal-1, infantile, 256730 600725 Holoprosencephaly-3, 142945 600759 Alzheimer disease-4 600760 Pseudohypoaldosteronism, type I, 264350 600760 Liddle syndrome, 177200 600761 Pseudohypoaldosteronism, type I, 264350 600761 Liddle syndrome, 177200 600795 Dementia, familial, nonspecific 600807 Bronchial asthma 600808 Enuresis, nocturnal, 2 600811 Xeroderma pigmentosum, group E, DDB-negative subtype, 278740 600837 Hirschsprung disease, 142623 600839 Bartter syndrome, 241200 600850 Schizophrenia disorder-4 600852 Retinitis pigmentosa-17 600856 Beckwith-Wiedemann syndrome, 130650 600881 Cataract, congenital, zonular, with sutural opacities 600882 Charcot-Marie-Tooth neuropathy-2B 600883 Diabetes mellitus, insulin-dependent, 8 600887 Endometrial carcinoma 600897 Cataract, zonular pulverulent-1, 116200 600900 Muscular dystrophy, limb-girdle, type 2E 600918 Cystinuria, type III 600923 Porphyria variegata, 176200 600937 Persistent hyperinsulinemic hypoglycemia of infancy, 256450 600946 Short stature, autosomal dominant, with normal serum growth hormone binding protein 600946 Short stature, idiopathic 600946 Laron dwarfism, 262500 600956 Persistent Mullerian duct syndrome, type II, 261550 600957 Persistent Mullerian duct syndrome, type I, 261550 600958 Cardiomyopathy, familial hypertrophic, 4, 115197 600965 Deafness, autosomal dominant 6 600968 Gitelman syndrome, 263800 600971 Deafness, autosomal recessive 6 600974 Deafness, autosomal recessive 7 600975 Glaucoma 3, primary infantile, B 600977 Cone dystrophy, progressive 600983 Pseudohypoaldosteronism type I, autosomal dominant, 177735 600993 Pancreatic cancer 600995 Nephrotic syndrome, idiopathic, steroid-resistant 600996 Arrhythmogenic right ventricular dysplasia-2 600998 Bleeding diathesis due to GNAQ deficiency 601002 5-oxoprolinuria, 266130 601002 Hemolytic anemia due to glutathione synthetase deficiency, 231900 601011 Spinocerebellar ataxia-6, 183086 601011 Cerebellar ataxia, pure 601011 Episodic ataxia, type 2, 108500 601011 Hemiplegic migraine, familial, 141500 601071 Deafness, autosomal recessive 9 601072 Deafness, autosomal recessive 8 601090 Iridogoniodysgenesis, 601631 601105 Pycnodysostosis, 265800 601107 Dubin-Johnson syndrome, 237500 601130 Tolbutamide poor metabolizer 601145 Epilepsy, progressive myoclonic 1, 254800 601146 Brachydactyly, type C, 113100 601146 Acromesomelic dysplasia, Hunter-Thompson type, 201250 601146 Chondrodysplasia, Grebe type, 200700 601154 Cardiomyopathy, dilated, 1E 601199 Neonatal hyperparathyroidism, 239200 601199 Hypocalcemia, autosomal dominant, 601198 601199 Hypocalciuric hypercalcemia, type I, 145980 601202 Cataract, anterior polar-2 601208 Insulin-dependent diabetes mellitus-11 601226 Progressive external ophthalmoplegia, type 2 601238 Cerebellar ataxia, Cayman type 601267 HIV infection, susceptibility/resistence to 601277 Ichthyosis, lamellar, type 2 601284 Hereditary hemorrhagic telangiectasia-2, 600376 601313 Polycystic kidney disease, adult type I, 173900 601316 Deafness, autosomal dominant 10 601318 Diabetes mellitus, insulin-dependent, 13 601362 DiGeorge syndrome/velocardiofacial syndrome complex-2 601363 Wilms tumor, type 4 601373 HIV infection, susceptibility/resistance to 601382 Charcot-Marie-Tooth neuropathy-4B 601385 Prostate cancer 601387 Breast cancer 601399 Platelet disorder, familial, with associated myeloid malignancy 601402 Leukemia, myeloid, acute 601406 B-cell non-Hodgkin lymphoma, high-grade 601410 Diabetes mellitus, transient neonatal 601411 Muscular dystrophy, limb-girdle, type 2F, 601287 601412 Deafness, autosomal dominant 7 601414 Retinitis pigmentosa-18 601458 Inflammatory bowel disease-2 601471 Moebius syndrome-2 601493 Cardiomyopathy, dilated 1C 601494 Cardiomyopathy, familial, dilated-2 601498 Peroxisomal biogenesis disorder, complementation group 4 601518 Prostate cancer, hereditary, 1, 176807 601545 Lissencephaly-1 601556 Spinocerebellar ataxia-1, 164400 601567 Combined factor V and VIII deficiency, 227300 601596 Charcot-Marie-Tooth neuropathy, demyelinating 601604 Mycobacterial and salmonella infections, susceptibility to 601606 Trichoepithelioma, multiple familial 601620 Holt-Oram syndrome, 142900 601621 Ulnar-mammary syndrome, 181450 601622 Saethre-Chotzen syndrome, 101400 601623 Angelman syndrome 601649 Blepharophimosis, epicanthus inversus, and ptosis, type 2 601650 Paraganglioma, familial nonchromaffin, 2 601652 Glaucoma 1A, primary open angle, juvenile-onset, 137750 601653 Branchiootic syndrome 601653 Branchiootorenal syndrome, 113650 601666 Insulin-dependent diabetes mellitus-15 601669 Hirschsprung disease, one form 601676 Acute insulin response 601680 Distal arthrogryposis, type 2B 601682 Glaucoma 1C, primary open angle 601687 Meesmann corneal dystrophy, 122100 601690 Platelet-activating factor acetylhydrolase deficiency 601691 Retinitis pigmentosa-19, 601718 601691 Stargardt disease-1, 248200 601691 Cone-rod dystrophy 3 601691 Fundus flavimaculatus with macular dystrophy, 248200 601692 Reis-Bucklers corneal dystrophy 601692 Corneal dystrophy, Avellino type 601692 Corneal dystrophy, Groenouw type I, 121900 601692 Corneal dystrophy, lattice type I, 122200 601718 Retinitis pigmentosa-19 601728 Bannayan-Zonana syndrome, 153480 601728 Cowden disease, 158350 601728 Endometrial carcinoma 601728 Lhermitte-Duclos syndrome 601744 Systemic lupus erythematosus, susceptibility to, 1 601757 Rhizomelic chondrodysplasia punctata, type 1, 215100 601768 Leukemia, acute myeloid 601769 Osteoporosis, involutional 601769 Rickets, vitamin D-resistant, 277440 601771 Glaucoma 3A, primary infantile, 231300 601777 Cone dystrophy, progressive 601780 Ceroid-lipofuscinosis, neuronal-6, variant late infantile 601785 Carbohydrate-deficient glycoprotein syndrome, type I, 212065 601800 [Hair color, brown] 601843 Hypothyroidism, congenital, 274400 601844 Pseudohypoaldosteronism type II 601846 Muscular dystrophy with rimmed vacuoles 601847 Progressive intrahepatic cholestasis-2 601850 Retinitis pigmentosa-deafness syndrome 601863 Bare lymphocyte syndrome, complementation group C 601868 Deafness, autosomal dominant 13 601884 [High bone mass] 601889 Lymphoma, diffuse large cell 601916 Pancreatic cancer 601928 Monilethrix, 158000 601941 Insulin-dependent diabetes mellitus-6 601954 Muscular dystrophy, limb-girdle, type 2G 601969 Medulloblastoma, 155255 601969 Glioblastoma multiforme, 137800 601975 Ectodermal dysplasia/skin fragility syndrome 601990 Neuroblastoma 602014 Hypomagnesemia with secondary hypocalcemia 602023 Bartter syndrome, type 3 602025 Obesity/hyperinsulinism, susceptibility to 602028 Multiple myeloma 602066 Convulsions, infantile and paroxysmal choreoathetosis 602067 Cardiomyopathy, dilated, 1F 602078 Fibrosis of extraocular muscles, congenital, 2 602080 Paget disease of bone-2 602081 Speech-language disorder-1 602082 Corneal dystrophy, Thiel-Behnke type 602084 Endometrial carcinoma 602085 Postaxial polydactyly, type A2 602086 Arrhythmogenic right ventricular dysplasia-3 602087 Arrhythmogenic right ventricular dysplasia-4 602088 Nephronophthisis, infantile 602089 Hemangioma, capillary, hereditary 602091 Marfan syndrome, atypical 602092 Deafness, autosomal recessive 18 602094 Lipodystrophy, familial partial 602096 Alzheimer disease-5 602099 Amytrophic lateral sclerosis-5 602116 Glioma 602117 Prader-Willi syndrome 602121 Deafness, autosomal dominant nonsyndromic sensorineural, 1, 124900 602134 Tremor, familial essential, 2 602136 Refsum disease, infantile, 266510 602136 Zellweger syndrome-1, 214100 602136 Adrenoleukodystrophy, neonatal, 202370 602153 Monilethrix, 158000 602216 Peutz-Jeghers syndrome, 175200 602225 Cone-rod retinal dystrophy-2, 120970 602225 Leber congenital amaurosis, type III 602235 Epilepsy, benign, neonatal, type 1, 121200 602279 Oculopharyngeal muscular dystorphy, 164300 602279 Oculopharyngeal muscular dystrophy, autosomal recessive, 257950 602280 Retinitis pigmentosa-14, 600132 602363 Ellis-van Creveld-like syndrome 602397 Cholestasis, benign recurrent intrahepatic, 243300 602397 Cholestasis, progressive familial intrahepatic-1, 211600 602403 Alzheimer disease, susceptibility to 602404 Parkinson disease, type 3 602447 Coronary artery disease, susceptibility to 602460 Deafness, autosomal dominant 15, 602459 602475 Ossification of posterior longitudinal ligament of spine 602476 Febrile convulsions, familial, 1 602477 Febrile convulsions, familial, 2 602491 Hyperlipidemia, familial combined, 1 602522 Bartter syndrome, infantile, with sensorineural deafness 602544 Parkinson disease, juvenile, type 2, 600116 602568 Homocystinuria-megaloblastic anemia, cbl E type, 236270 602574 Deafness, autosomal dominant 12, 601842 602574 Deafness, autosomal dominant 8, 601543 602575 Nail-patella syndrome with open-angle glaucoma, 137750 602575 Nail-patella syndrome, 161200 602616 Carbohydrate-deficient glycoprotein syndrome, type II, 212066 602629 Dystonia-6, torsion 602631 Rhabdomyosarcoma, 268210 602631 Breast Cancer 602669 Anterior segment mesenchymal dysgenesis and cataract, 107250 602669 Cataract, congenital 602685 Mental retardation, severe, with spasticity and tapetoretinal degeneration 602716 Nephrosis-1, congenital, Finnish type, 256300 602759 Prostate cancer, hereditary, 2, 176807 602771 Muscular dystrophy, congenital, with early spine rigidity 602772 Retinitis pitmentosa-24 602782 Faisalabad histiocytosis 602783 Spastic paraplegia-7 Mature Polypeptides

The present invention also encompasses mature forms of a polypeptide having the amino acid sequence of SEQ ID NO:Y and/or the amino acid sequence encoded by the cDNA in a deposited clone. Polynucleotides encoding the mature forms (such as, for example, the polynucleotide sequence in SEQ ID NO:X and/or the polynucleotide sequence contained in the cDNA of a deposited clone) are also encompassed by the invention. Moreover, fragments or variants of these polypeptides (such as, fragments as described herein, polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to these polypeptides, or polypeptides encoded by a polynucleotide that hybridizes under stringent conditions to the complementary strand of the polynucleotide encoding these polypeptides) are also encompassed by the invention. In preferred embodiments, these fragments or variants retain one or more functional activities of the full-length or mature form of the polypeptide (e.g., biological activity (such as, for example, activity in detecting, preventing, treating and/or indicated disorders), antigenicity (ability to bind, or compete with a polypeptide of the invention for binding, to an anti-polypeptide of the invention antibody), immunogenicity (ability to generate antibody which binds to a specific polypeptide of the invention), ability to form multimers with polypeptides of the invention, and ability to bind to a receptor or ligand for a polypeptide of the invention). Antibodies that bind the polypeptides of the invention, and polynucleotides encoding these polypeptides are also encompassed by the invention.

According to the signal hypothesis, proteins secreted by mammalian cells have a signal or secretary leader sequence which is cleaved from the mature protein once export of the growing protein chain across the rough endoplasmic reticulum has been initiated. Most mammalian cells and even insect cells cleave secreted proteins with the same specificity. However, in some cases, cleavage of a secreted protein is not entirely uniform, which results in two or more mature species of the protein. Further, it has long been known that cleavage specificity of a secreted protein is ultimately determined by the primary structure of the complete protein, that is, it is inherent in the amino acid sequence of the polypeptide.

Methods for predicting whether a protein has a signal sequence, as well as the cleavage point for that sequence, are available. For instance, the method of McGeoch, Virus Res. 3:271-286 (1985), uses the information from a short N-terminal charged region and a subsequent uncharged region of the complete (uncleaved) protein. The method of von Heinje, Nucleic Acids Res. 14:4683-4690 (1986) uses the information from the residues surrounding the cleavage site, typically residues −13 to +2, where +1 indicates the amino terminus of the secreted protein. The accuracy of predicting the cleavage points of known mammalian secretory proteins for each of these methods is in the range of 75-80%. (von Heinje, supra.) However, the two methods do not always produce the same predicted cleavage point(s) for a given protein.

In the present case, the deduced amino acid sequence of the secreted polypeptide was analyzed by a computer program called SignalP (Henrik Nielsen et al., Protein Engineering 10:1-6 (1997)), which predicts the cellular location of a protein based on the amino acid sequence. As part of this computational prediction of localization, the methods of McGeoch and von Heinje are incorporated. The analysis of the amino acid sequences of the secreted proteins described herein by this program provided the results shown in Table 1A.

In specific embodiments, polypeptides of the invention comprise, or alternatively consist of, the predicted mature form of the polypeptide as delineated in columns 14 and 15 of Table 1A. Moreover, fragments or variants of these polypeptides (such as, fragments as described herein, polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to these polypeptides, or polypeptides encoded by a polynucleotide that hybridizes under stringent conditions to the complementary strand of the polynucleotide encoding these polypeptides) are also encompassed by the invention. In preferred embodiments, these fragments or variants retain one or more functional activities of the full-length or mature form of the polypeptide (e.g., biological activity, antigenicity [ability to bind (or compete with a polypeptide of the invention for binding) to an anti-polypeptide of the invention antibody], immunogenicity (ability to generate antibody which binds to a specific polypeptide of the invention), ability to form multimers with polypeptides of the invention, and ability to bind to a receptor or ligand for a polypeptide of the invention). Antibodies that bind the polypeptides of the invention, and polynucleotides encoding these polypeptides are also encompassed by the invention.

Polynucleotides encoding proteins comprising, or consisting of, the predicted mature form of polypeptides of the invention (e.g., polynucleotides having the sequence of SEQ ID NO: X (Table 1A, column 4), the sequence delineated in columns 7 and 8 of Table 1A, and a sequence encoding the mature polypeptide delineated in columns 14 and 15 of Table 1A (e.g., the sequence of SEQ ID NO:X encoding the mature polypeptide delineated in columns 14 and 15 of Table 1)) are also encompassed by the invention, as are fragments or variants of these polynucleotides (such as, fragments as described herein, polynucleotides at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to these polynucleotides, and nucleic acids which hybridizes under stringent conditions to the complementary strand of the polynucleotide).

As one of ordinary skill would appreciate, however, cleavage sites sometimes vary from organism to organism and cannot be predicted with absolute certainty. Accordingly, the present invention provides secreted polypeptides having a sequence shown in SEQ ID NO:Y which have an N-terminus beginning within 15 residues of the predicted cleavage point (i.e., having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 more or less contiguous residues of SEQ ID NO:Y at the N-terminus when compared to the predicted mature form of the polypeptide (e.g., the mature polypeptide delineated in columns 14 and 15 of Table 1). Similarly, it is also recognized that in some cases, cleavage of the signal sequence from a secreted protein is not entirely uniform, resulting in more than one secreted species. These polypeptides, and the polynucleotides encoding such polypeptides, are contemplated by the present invention.

Moreover, the signal sequence identified by the above analysis may not necessarily predict the naturally occurring signal sequence. For example, the naturally occurring signal sequence may be further upstream from the predicted signal sequence. However, it is likely that the predicted signal sequence will be capable of directing the secreted protein to the ER. Nonetheless, the present invention provides the mature protein produced by expression of the polynucleotide sequence of SEQ ID NO:X and/or the polynucleotide sequence contained in the cDNA of a deposited clone, in a mammalian cell (e.g., COS cells, as described below). These polypeptides, and the polynucleotides encoding such polypeptides, are contemplated by the present invention.

Polynucleotide and Polypeptide Variants

The present invention is also directed to variants of the polynucleotide sequence disclosed in SEQ ID NO:X or the complementary strand thereto, nucleotide sequences encoding the polypeptide of SEQ ID NO:Y, the nucleotide sequence of SEQ ID NO:X that encodes the polypeptide sequence as defined in columns 13 and 14 of Table 1A, nucleotide sequences encoding the polypeptide sequence as defined in columns 13 and 14 of Table 1A, the nucleotide sequence of SEQ ID NO:X encoding the polypeptide sequence as defined in column 5 of Table 1B.1, nucleotide sequences encoding the polypeptide as defined in column 6 and column 7 of Table 1B.1, the nucleotide sequence as defined in columns 8 and 9 of Table 2, nucleotide sequences encoding the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2, the nucleotide sequence as defined in column 6 of Table 1C, nucleotide sequences encoding the polypeptide encoded by the nucleotide sequence as defined in column 6 of Table 1C, the cDNA sequence contained in ATCC Deposit NO:Z, nucleotide sequences encoding the polypeptide encoded by the cDNA sequence contained in ATCC Deposit NO:Z, and/or nucleotide sequences encoding a mature (secreted) polypeptide encoded by the cDNA sequence contained in ATCC Deposit NO:Z.

The present invention also encompasses variants of the polypeptide sequence disclosed in SEQ ID NO:Y, the polypeptide as defined in columns 13 and 14 of Table 1A, the polypeptide sequence as defined in columns 6 and 7 of Table 1B.1, a polypeptide sequence encoded by the polynucleotide sequence in SEQ ID NO:X, a polypeptide sequence encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2, a polypeptide sequence encoded by the nucleotide sequence as defined in column 6 of Table 1C, a polypeptide sequence encoded by the complement of the polynucleotide sequence in SEQ ID NO:X, the polypeptide sequence encoded by the cDNA sequence contained in ATCC Deposit NO:Z and/or a mature (secreted) polypeptide encoded by the cDNA sequence contained in ATCC Deposit NO:Z.

“Variant” refers to a polynucleotide or polypeptide differing from the polynucleotide or polypeptide of the present invention, but retaining essential properties thereof. Generally, variants are overall closely similar, and, in many regions, identical to the polynucleotide or polypeptide of the present invention.

Thus, one aspect of the invention provides an isolated nucleic acid molecule comprising, or alternatively consisting of, a polynucleotide having a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence described in SEQ ID NO:X or contained in the cDNA sequence of ATCC Deposit No:Z; (b) a nucleotide sequence in SEQ ID NO:X or the cDNA in ATCC Deposit No:Z which encodes the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in ATCC Deposit No:Z; (c) a nucleotide sequence in SEQ ID NO:X or the cDNA in ATCC Deposit No:Z which encodes a mature polypeptide (i.e., a secreted polypeptide (e.g., as delineated in columns 14 and 15 of Table 1A)); (d) a nucleotide sequence in SEQ ID NO:X or the cDNA sequence of ATCC Deposit No:Z, which encodes a biologically active fragment of a polypeptide; (e) a nucleotide sequence in SEQ ID NO:X or the cDNA sequence of ATCC Deposit No:Z, which encodes an antigenic fragment of a polypeptide; (f) a nucleotide sequence encoding a polypeptide comprising the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in ATCC Deposit No:Z; (g) a nucleotide sequence encoding a mature polypeptide of the amino acid sequence of SEQ ID NO:Y (i.e., a secreted polypeptide (e.g., as delineated in columns 14 and 15 of Table 1A)) or a mature polypeptide of the amino acid sequence encoded by the cDNA in ATCC Deposit No:Z; (h) a nucleotide sequence encoding a biologically active fragment of a polypeptide having the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in ATCC Deposit No:Z; (i) a nucleotide sequence encoding an antigenic fragment of a polypeptide having the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in ATCC Deposit No:Z; and (j) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), (h), or (i) above.

The present invention is also directed to nucleic acid molecules which comprise, or alternatively consist of, a nucleotide sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100%, identical to, for example, any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), (h), (i), or (j) above, the nucleotide coding sequence in SEQ ID NO:X or the complementary strand thereto, the nucleotide coding sequence of the cDNA contained in ATCC Deposit No:Z or the complementary strand thereto, a nucleotide sequence encoding the polypeptide of SEQ ID NO:Y, a nucleotide sequence encoding a polypeptide sequence encoded by the nucleotide sequence in SEQ ID NO:X, a polypeptide sequence encoded by the complement of the polynucleotide sequence in SEQ ID NO:X, a nucleotide sequence encoding the polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, the nucleotide coding sequence in SEQ ID NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto, a nucleotide sequence encoding the polypeptide encoded by the nucleotide sequence in SEQ ID NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto, the nucleotide coding sequence in SEQ ID NO:B as defined in column 6 of Table 1C or the complementary strand thereto, a nucleotide sequence encoding the polypeptide encoded by the nucleotide sequence in SEQ ID NO:B as defined in column 6 of Table 1C or the complementary strand thereto, the nucleotide sequence in SEQ ID NO:X encoding the polypeptide sequence as defined in columns 6 and 7 of Table 1B.1 or the complementary strand thereto, nucleotide sequences encoding the polypeptide as defined in column 6 and 7 of Table 1B.1 or the complementary strand thereto, and/or polynucleotide fragments of any of these nucleic acid molecules (e.g., those fragments described herein). Polynucleotides which hybridize to the complement of these nucleic acid molecules under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention, as are polypeptides encoded by these polynucleotides and nucleic acids.

In a preferred embodiment, the invention encompasses nucleic acid molecules which comprise, or alternatively, consist of a polynucleotide which hybridizes under stringent hybridization conditions, or alternatively, under lower stringency conditions, to a polynucleotide in (a), (b), (c), (d), (e), (f), (g), (h), or (i), above, as are polypeptides encoded by these polynucleotides. In another preferred embodiment, polynucleotides which hybridize to the complement of these nucleic acid molecules under stringent hybridization conditions, or alternatively, under lower stringency conditions, are also encompassed by the invention, as are polypeptides encoded by these polynucleotides.

In another embodiment, the invention provides a purified protein comprising, or alternatively consisting of, a polypeptide having an amino acid sequence selected from the group consisting of: (a) the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in ATCC Deposit No:Z; (b) the amino acid sequence of a mature (secreted) form of a polypeptide having the amino acid sequence of SEQ ID NO:Y (e.g., as delineated in columns 14 and 15 of Table 1A) or a mature form of the amino acid sequence encoded by the cDNA in ATCC Deposit No:Z mature; (c) the amino acid sequence of a biologically active fragment of a polypeptide having the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in ATCC Deposit No:Z; and (d) the amino acid sequence of an antigenic fragment of a polypeptide having the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in ATCC Deposit No:Z.

The present invention is also directed to proteins which comprise, or alternatively consist of, an amino acid sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100%, identical to, for example, any of the amino acid sequences in (a), (b), (c), or (d), above, the amino acid sequence shown in SEQ ID NO:Y, the amino acid sequence encoded by the cDNA contained in ATCC Deposit No:Z, the amino acid sequence of the polypeptide encoded by the nucleotide sequence in SEQ ID NO:X as defined in columns 8 and 9 of Table 2, the amino acid sequence of the polypeptide encoded by the nucleotide sequence in SEQ ID NO:B as defined in column 6 of Table 1C, the amino acid sequence as defined in column 6 and 7 of Table 1B.1, an amino acid sequence encoded by the nucleotide sequence in SEQ ID NO:X, and an amino acid sequence encoded by the complement of the polynucleotide sequence in SEQ ID NO:X. Fragments of these polypeptides are also provided (e.g., those fragments described herein). Further proteins encoded by polynucleotides which hybridize to the complement of the nucleic acid molecules encoding these amino acid sequences under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention, as are the polynucleotides encoding these proteins.

By a nucleic acid having a nucleotide sequence at least, for example, 95% “identical” to a reference nucleotide sequence of the present invention, it is intended that the nucleotide sequence of the nucleic acid is identical to the reference sequence except that the nucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding the polypeptide. In other words, to obtain a nucleic acid having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. The query sequence may be an entire sequence referred to in Table 1B or 2 as the ORF (open reading frame), or any fragment specified as described herein.

As a practical matter, whether any particular nucleic acid molecule or polypeptide is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleotide sequence of the present invention can be determined conventionally using known computer programs. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990)). In a sequence alignment the query and subject sequences are both DNA sequences. An RNA sequence can be compared by converting U's to T's. The result of said global sequence alignment is expressed as percent identity. Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identity are: Matrix=Unitary, k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject nucleotide sequence, whichever is shorter.

If the subject sequence is shorter than the query sequence because of 5′ or 3′ deletions, not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB program does not account for 5′ and 3′ truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5′ or 3′ ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are 5′ and 3′ of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence. Whether a nucleotide is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score is what is used for the purposes of the present invention. Only bases outside the 5′ and 3′ bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes of manually adjusting the percent identity score.

For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5′ end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5′ end. The 10 unpaired bases represent 10% of the sequence (number of bases at the 5′ and 3′ ends not matched/total number of bases in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5′ or 3′ of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only bases 5′ and 3′ of the subject sequence which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are to be made for the purposes of the present invention.

By a polypeptide having an amino acid sequence at least, for example, 95% “identical” to a query amino acid sequence of the present invention, it is intended that the amino acid sequence of the subject polypeptide is identical to the query sequence except that the subject polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the query amino acid sequence. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a query amino acid sequence, up to 5% of the amino acid residues in the subject sequence may be inserted, deleted, (indels) or substituted with another amino acid. These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.

As a practical matter, whether any particular polypeptide is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the amino acid sequence of a polypeptide referred to in Table 1A (e.g., the amino acid sequence delineated in columns 14 and 15) or a fragment thereof, Table 1B.1 (e.g., the amino acid sequence identified in column 6) or a fragment thereof, Table 2 (e.g., the amino acid sequence of the polypeptide encoded by the polynucleotide sequence defined in columns 8 and 9 of Table 2) or a fragment thereof, the amino acid sequence of the polypeptide encoded by the polynucleotide sequence in SEQ ID NO:B as defined in column 6 of Table 1C or a fragment thereof, the amino acid sequence of the polypeptide encoded by the nucleotide sequence in SEQ ID NO:X or a fragment thereof, or the amino acid sequence of the polypeptide encoded by cDNA contained in ATCC Deposit No:Z, or a fragment thereof, the amino acid sequence of a mature (secreted) polypeptide encoded by cDNA contained in ATCC Deposit No:Z, or a fragment thereof, can be determined conventionally using known computer programs. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990)). In a sequence alignment the query and subject sequences are either both nucleotide sequences or both amino acid sequences. The result of said global sequence alignment is expressed as percent identity. Preferred parameters used in a FASTDB amino acid alignment are: Matrix=PAM 0, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1, Window Size=sequence length, Gap Penalty=5, Gap Size Penalty=0.05, Window Size=500 or the length of the subject amino acid sequence, whichever is shorter.

If the subject sequence is shorter than the query sequence due to N- or C-terminal deletions, not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB program does not account for N- and C-terminal truncations of the subject sequence when calculating global percent identity. For subject sequences truncated at the N- and C-termini, relative to the query sequence, the percent identity is corrected by calculating the number of residues of the query sequence that are N- and C-terminal of the subject sequence, which are not matched/aligned with a corresponding subject residue, as a percent of the total bases of the query sequence. Whether a residue is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This final percent identity score is what is used for the purposes of the present invention. Only residues to the N- and C-termini of the subject sequence, which are not matched/aligned with the query sequence, are considered for the purposes of manually adjusting the percent identity score. That is, only query residue positions outside the farthest N- and C-terminal residues of the subject sequence.

For example, a 90 amino acid residue subject sequence is aligned with a 100 residue query sequence to determine percent identity. The deletion occurs at the N-terminus of the subject sequence and therefore, the FASTDB alignment does not show a matching/alignment of the first 10 residues at the N-terminus. The 10 unpaired residues represent 10% of the sequence (number of residues at the N- and C-termini not matched/total number of residues in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 residues were perfectly matched the final percent identity would be 90%. In another example, a 90 residue subject sequence is compared with a 100 residue query sequence. This time the deletions are internal deletions so there are no residues at the N- or C-termini of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only residue positions outside the N- and C-terminal ends of the subject sequence, as displayed in the FASTDB alignment, which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are to made for the purposes of the present invention.

The polynucleotide variants of the invention may contain alterations in the coding regions, non-coding regions, or both. Especially preferred are polynucleotide variants containing alterations which produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide. Nucleotide variants produced by silent substitutions due to the degeneracy of the genetic code are preferred. Moreover, polypeptide variants in which less than 50, less than 40, less than 30, less than 20, less than 10, or 5-50, 5-25, 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination are also preferred. Polynucleotide variants can be produced for a variety of reasons, e.g., to optimize codon expression for a particular host (change codons in the human mRNA to those preferred by a bacterial host such as E. coli).

Naturally occurring variants are called “allelic variants,” and refer to one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. (Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985)). These allelic variants can vary at either the polynucleotide and/or polypeptide level and are included in the present invention. Alternatively, non-naturally occurring variants may be produced by mutagenesis techniques or by direct synthesis.

Using known methods of protein engineering and recombinant DNA technology, variants may be generated to improve or alter the characteristics of the polypeptides of the present invention. For instance, one or more amino acids can be deleted from the N-terminus or C-terminus of the polypeptide of the present invention without substantial loss of biological function. As an example, Ron et al. (J. Biol. Chem. 268: 2984-2988 (1993)) reported variant KGF proteins having heparin binding activity even after deleting 3, 8, or 27 amino-terminal amino acid residues. Similarly, Interferon gamma exhibited up to ten times higher activity after deleting 8-10 amino acid residues from the carboxy terminus of this protein. (Dobeli et al., J. Biotechnology 7:199-216 (1988).)

Moreover, ample evidence demonstrates that variants often retain a biological activity similar to that of the naturally occurring protein. For example, Gayle and coworkers (J. Biol. Chem. 268:22105-22111 (1993)) conducted extensive mutational analysis of human cytokine IL-1a. They used random mutagenesis to generate over 3,500 individual IL-1a mutants that averaged 2.5 amino acid changes per variant over the entire length of the molecule. Multiple mutations were examined at every possible amino acid position. The investigators found that “[m]ost of the molecule could be altered with little effect on either [binding or biological activity].” In fact, only 23 unique amino acid sequences, out of more than 3,500 nucleotide sequences examined, produced a protein that significantly differed in activity from wild-type.

Furthermore, even if deleting one or more amino acids from the N-terminus or C-terminus of a polypeptide results in modification or loss of one or more biological functions, other biological activities may still be retained. For example, the ability of a deletion variant to induce and/or to bind antibodies which recognize the secreted form will likely be retained when less than the majority of the residues of the secreted form are removed from the N-terminus or C-terminus. Whether a particular polypeptide lacking N- or C-terminal residues of a protein retains such immunogenic activities can readily be determined by routine methods described herein and otherwise known in the art.

Thus, the invention further includes polypeptide variants which show a functional activity (e.g., biological activity) of the polypeptides of the invention. Such variants include deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as have little effect on activity.

The present application is directed to nucleic acid molecules at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequences disclosed herein, (e.g., encoding a polypeptide having the amino acid sequence of an N and/or C terminal deletion), irrespective of whether they encode a polypeptide having functional activity. This is because even where a particular nucleic acid molecule does not encode a polypeptide having functional activity, one of skill in the art would still know how to use the nucleic acid molecule, for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer. Uses of the nucleic acid molecules of the present invention that do not encode a polypeptide having functional activity include, inter alia, (1) isolating a gene or allelic or splice variants thereof in a cDNA library; (2) in situ hybridization (e.g., “FISH”) to metaphase chromosomal spreads to provide precise chromosomal location of the gene, as described in Verma et al., Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York (1988); (3) Northern Blot analysis for detecting mRNA expression in specific tissues (e.g., normal or diseased tissues); and (4) in situ hybridization (e.g., histochemistry) for detecting mRNA expression in specific tissues (e.g., normal or diseased tissues).

Preferred, however, are nucleic acid molecules having sequences at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequences disclosed herein, which do, in fact, encode a polypeptide having functional activity. By a polypeptide having “functional activity” is meant, a polypeptide capable of displaying one or more known functional activities associated with a full-length (complete) protein and/or a mature (secreted) protein of the invention. Such functional activities include, but are not limited to, biological activity, antigenicity [ability to bind (or compete with a polypeptide of the invention for binding) to an anti-polypeptide of the invention antibody], immunogenicity (ability to generate antibody which binds to a specific polypeptide of the invention), ability to form multimers with polypeptides of the invention, and ability to bind to a receptor or ligand for a polypeptide of the invention.

The functional activity of the polypeptides, and fragments, variants and derivatives of the invention, can be assayed by various methods.

For example, in one embodiment where one is assaying for the ability to bind or compete with a full-length polypeptide of the present invention for binding to an anti-polypeptide antibody, various immunoassays known in the art can be used, including but not limited to, competitive and non-competitive assay systems using techniques such as radioimmunoassays, ELISA (enzyme linked immunosorbent assay), “sandwich” immunoassays, immunoradiometric assays, gel diffusion precipitation reactions, immunodiffusion assays, in situ immunoassays (using colloidal gold, enzyme or radioisotope labels, for example), western blots, precipitation reactions, agglutination assays (e.g., gel agglutination assays, hemagglutination assays), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc. In one embodiment, antibody binding is detected by detecting a label on the primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labeled. Many means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention.

In another embodiment, where a ligand is identified, or the ability of a polypeptide fragment, variant or derivative of the invention to multimerize is being evaluated, binding can be assayed, e.g., by means well-known in the art, such as, for example, reducing and non-reducing gel chromatography, protein affinity chromatography, and affinity blotting. See generally, Phizicky et al., Microbiol. Rev. 59:94-123 (1995). In another embodiment, the ability of physiological correlates of a polypeptide of the present invention to bind to a substrate(s) of the polypeptide of the invention can be routinely assayed using techniques known in the art.

In addition, assays described herein (see Examples) and otherwise known in the art may routinely be applied to measure the ability of polypeptides of the present invention and fragments, variants and derivatives thereof to elicit polypeptide related biological activity (either in vitro or in vivo). Other methods will be known to the skilled artisan and are within the scope of the invention.

Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of the nucleic acid molecules having a sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to, for example, the nucleic acid sequence of the cDNA contained in ATCC Deposit No:Z, the nucleic acid sequence referred to in Table 1B (SEQ ID NO:X), the nucleic acid sequence disclosed in Table 1A (e.g., the nucleic acid sequence delineated in columns 7 and 8), the nucleic acid sequence disclosed in Table 2 (e.g., the nucleic acid sequence delineated in columns 8 and 9) or fragments thereof, will encode polypeptides “having functional activity.” In fact, since degenerate variants of any of these nucleotide sequences all encode the same polypeptide, in many instances, this will be clear to the skilled artisan even without performing the above described comparison assay. It will be further recognized in the art that, for such nucleic acid molecules that are not degenerate variants, a reasonable number will also encode a polypeptide having functional activity. This is because the skilled artisan is fully aware of amino acid substitutions that are either less likely or not likely to significantly effect protein function (e.g., replacing one aliphatic amino acid with a second aliphatic amino acid), as further described below.

For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in Bowie et al., “Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions,” Science 247:1306-1310 (1990), wherein the authors indicate that there are two main strategies for studying the tolerance of an amino acid sequence to change.

The first strategy exploits the tolerance of amino acid substitutions by natural selection during the process of evolution. By comparing amino acid sequences in different species, conserved amino acids can be identified. These conserved amino acids are likely important for protein function. In contrast, the amino acid positions where substitutions have been tolerated by natural selection indicates that these positions are not critical for protein function. Thus, positions tolerating amino acid substitution could be modified while still maintaining biological activity of the protein.

The second strategy uses genetic engineering to introduce amino acid changes at specific positions of a cloned gene to identify regions critical for protein function. For example, site directed mutagenesis or alanine-scanning mutagenesis (introduction of single alanine mutations at every residue in the molecule) can be used. See Cunningham and Wells, Science 244:1081-1085 (1989). The resulting mutant molecules can then be tested for biological activity.

As the authors state, these two strategies have revealed that proteins are surprisingly tolerant of amino acid substitutions. The authors further indicate which amino acid changes are likely to be permissive at certain amino acid positions in the protein. For example, most buried (within the tertiary structure of the protein) amino acid residues require nonpolar side chains, whereas few features of surface side chains are generally conserved. Moreover, tolerated conservative amino acid substitutions involve replacement of the aliphatic or hydrophobic amino acids Ala, Val, Leu and lie; replacement of the hydroxyl residues Ser and Thr; replacement of the acidic residues Asp and Glu; replacement of the amide residues Asn and Gin, replacement of the basic residues Lys, Arg, and His; replacement of the aromatic residues Phe, Tyr, and Trp, and replacement of the small-sized amino acids Ala, Ser, Thr, Met, and Gly.

Besides conservative amino acid substitution, variants of the present invention include (i) substitutions with one or more of the non-conserved amino acid residues, where the substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) substitutions with one or more of the amino acid residues having a substituent group, or (iii) fusion of the mature polypeptide with another compound, such as a compound to increase the stability and/or solubility of the polypeptide (for example, polyethylene glycol), (iv) fusion of the polypeptide with additional amino acids, such as, for example, an IgG Fc fusion region peptide, serum albumin (preferably human serum albumin) or a fragment thereof, or leader or secretory sequence, or a sequence facilitating purification, or (v) fusion of the polypeptide with another compound, such as albumin (including but not limited to recombinant albumin (see, e.g., U.S. Pat. No. 5,876,969, issued Mar. 2, 1999, EP Patent 0 413 622, and U.S. Pat. No. 5,766,883, issued Jun. 16, 1998, herein incorporated by reference in their entirety)). Such variant polypeptides are deemed to be within the scope of those skilled in the art from the teachings herein.

For example, polypeptide variant containing amino acid substitutions of charged amino acids with other charged or neutral amino acids may produce proteins with improved characteristics, such as less aggregation. Aggregation of pharmaceutical formulations both reduces activity and increases clearance due to the aggregate's immunogenic activity. See Pinckard et al., Clin. Exp. Immunol. 2:331-340 (1967); Robbins et al., Diabetes 36: 838-845 (1987); Cleland et al., Crit. Rev. Therapeutic Drug Carrier Systems 10:307-377 (1993).

A further embodiment of the invention relates to polypeptides which comprise the amino acid sequence of a polypeptide having an amino acid sequence which contains at least one amino acid substitution, but not more than 50 amino acid substitutions, even more preferably, not more than 40 amino acid substitutions, still more preferably, not more than 30 amino acid substitutions, and still even more preferably, not more than 20 amino acid substitutions from a polypeptide sequence disclosed herein. Of course it is highly preferable for a polypeptide to have an amino acid sequence which, for example, comprises the amino acid sequence of a polypeptide of SEQ ID NO:Y, the amino acid sequence of the mature (e.g., secreted) polypeptide of SEQ ID NO:Y, an amino acid sequence encoded by SEQ ID NO:X, an amino acid sequence encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, an amino acid sequence encoded by the complement of SEQ ID NO:X, an amino acid sequence encoded by cDNA contained in ATCC Deposit No:Z, and/or the amino acid sequence of a mature (secreted) polypeptide encoded by cDNA contained in ATCC Deposit No:Z, or a fragment thereof, which contains, in order of ever-increasing preference, at least one, but not more than 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitutions.

In specific embodiments, the polypeptides of the invention comprise, or alternatively, consist of, fragments or variants of a reference amino acid sequence selected from: (a) the amino acid sequence of SEQ ID NO:Y or fragments thereof (e.g., the mature form and/or other fragments described herein); (b) the amino acid sequence encoded by SEQ ID NO:X or fragments thereof; (c) the amino acid sequence encoded by the complement of SEQ ID NO:X or fragments thereof; (d) the amino acid sequence encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2 or fragments thereof; and (e) the amino acid sequence encoded by cDNA contained in ATCC Deposit No:Z or fragments thereof; wherein the fragments or variants have 1-5, 5-10, 5-25, 5-50, 10-50 or 50-150, amino acid residue additions, substitutions, and/or deletions when compared to the reference amino acid sequence. In preferred embodiments, the amino acid substitutions are conservative. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Polynucleotide and Polypeptide Fragments

The present invention is also directed to polynucleotide fragments of the polynucleotides (nucleic acids) of the invention. In the present invention, a “polynucleotide fragment” refers to a polynucleotide having a nucleic acid sequence which, for example: is a portion of the cDNA contained in ATCC Deposit No:Z or the complementary strand thereto; is a portion of the polynucleotide sequence encoding the polypeptide encoded by the cDNA contained in ATCC Deposit No:Z or the complementary strand thereto; is a portion of the polynucleotide sequence encoding the mature (secreted) polypeptide encoded by the cDNA contained in ATCC Deposit No:Z or the complementary strand thereto; is a portion of a polynucleotide sequence encoding the mature amino acid sequence as defined in columns 14 and 15 of Table 1A or the complementary strand thereto; is a portion of a polynucleotide sequence encoding the amino acid sequence encoded by the region of SEQ ID NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto; is a portion of the polynucleotide sequence of SEQ ID NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto; is a portion of the polynucleotide sequence in SEQ ID NO:X or the complementary strand thereto; is a polynucleotide sequence encoding a portion of the polypeptide of SEQ ID NO:Y; is a polynucleotide sequence encoding a portion of a polypeptide encoded by SEQ ID NO:X; is a polynucleotide sequence encoding a portion of a polypeptide encoded by the complement of the polynucleotide sequence in SEQ ID NO:X; is a portion of a polynucleotide sequence encoding the amino acid sequence encoded by the region of SEQ ID NO:B as defined in column 6 of Table 1C or the complementary strand thereto; or is a portion of the polynucleotide sequence of SEQ ID NO:B as defined in column 6 of Table 1C or the complementary strand thereto.

The polynucleotide fragments of the invention are preferably at least about 15 nt, and more preferably at least about 20 nt, still more preferably at least about 30 nt, and even more preferably, at least about 40 nt, at least about 50 nt, at least about 75 nt, or at least about 150 nt in length. A fragment “at least 20 nt in length,” for example, is intended to include 20 or more contiguous bases from the cDNA sequence contained in ATCC Deposit No:Z, or the nucleotide sequence shown in SEQ ID NO:X or the complementary stand thereto. In this context “about” includes the particularly recited value or a value larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. These nucleotide fragments have uses that include, but are not limited to, as diagnostic probes and primers as discussed herein. Of course, larger fragments (e.g., at least 160, 170, 180, 190, 200, 250, 500, 600, 1000, or 2000 nucleotides in length) are also encompassed by the invention.

Moreover, representative examples of polynucleotide fragments of the invention comprise, or alternatively consist of, a sequence from about nucleotide number 1-50, 51-100, 101-150, 151-200, 201-250, 251-300, 301-350, 351-400, 401-450, 451-500, 501-550, 551-600, 601-650, 651-700, 701-750, 751-800, 801-850, 851-900, 901-950, 951-1000, 1001-1050, 1051-1100, 1101-1150, 1151-1200, 1201-1250, 1251-1300, 1301-1350, 1351-1400, 1401-1450, 1451-1500, 1501-1550, 1551-1600, 1601-1650, 1651-1700, 1701-1750, 1751-1800, 1801-1850, 1851-1900, 1901-1950, 1951-2000, 2001-2050, 2051-2100, 2101-2150, 2151-2200, 2201-2250, 2251-2300, 2301-2350, 2351-2400, 2401-2450, 2451-2500, 2501-2550, 2551-2600, 2601-2650, 2651-2700, 2701-2750, 2751-2800, 2801-2850, 2851-2900, 2901-2950, 2951-3000, 3001-3050, 3051-3100, 3101-3150, 3151-3200, 3201-3250, 3251-3300, 3301-3350, 3351-3400, 3401-3450, 3451-3500, 3501-3550, 3551-3600, 3601-3650, 3651-3700, 3701-3750, 3751-3800, 3801-3850, 3851-3900, 3901-3950, 3951-4000, 4001-4050, 4051-4100, 4101-4150, 4151-4200, 4201-4250, 4251-4300, 4301-4350, 4351-4400, 4401-4450, 4451-4500, 4501-4550, 4551-4600, 4601-4650, 4651-4700, 4701-4750, 4751-4800, 4801-4850, 4851-4900, 4901-4950, 4951-5000, 5001-5050, 5051-5100, 5101-5150, 5151-5200, 5201-5250, 5251-5300, 5301-5350, 5351-5400, 5401-5450, 5451-5500, 5501-5550, 5551-5600, 5601-5650, 5651-5700, 5701-5750, 5751-5800, 5801-5850, 5851-5900, 5901-5950, 5951-6000, 6001-6050, 6051-6100, 6101-6150, 6151-6200, 6201-6250, 6251-6300, 6301-6350, 6351-6400, 6401-6450, 6451-6500, 6501-6550, 6551-6600, 6601-6650, 6651-6700, 6701-6750, 6751-6800, 6801-6850, 6851-6900, 6901-6950, 6951-7000, 7001-7050, 7051-7100, 7101-7150, 7151-7200, 7201-7250, 7251-7300 or 7301 to the end of SEQ ID NO:X, or the complementary strand thereto. In this context “about” includes the particularly recited range or a range larger or smaller by several (5, 4, 3, 2, or t) nucleotides, at either terminus or at both termini. Preferably, these fragments encode a polypeptide which has a functional activity (e.g., biological activity). More preferably, these polynucleotides can be used as probes or primers as discussed herein. Polynucleotides which hybridize to one or more of these polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions are also encompassed by the invention, as are polypeptides encoded by these polynucleotides.

Further representative examples of polynucleotide fragments of the invention comprise, or alternatively consist of, a sequence from about nucleotide number 1-50, 51-100, 101-150, 151-200, 201-250, 251-300, 301-350, 351-400, 401-450, 451-500, 501-550, 551-600, 601-650, 651-700, 701-750, 751-800, 801-850, 851-900, 901-950, 951-1000, 1001-1050, 1051-1100, 1101-1150, 1151-1200, 1201-1250, 1251-1300, 1301-1350, 1351-1400, 1401-1450, 1451-1500, 1501-1550, 1551-1600, 1601-1650, 1651-1700, 1701-1750, 1751-1800, 1801-1850, 1851-1900, 1901-1950, 1951-2000, 2001-2050, 2051-2100, 2101-2150, 2151-2200, 2201-2250, 2251-2300, 2301-2350, 2351-2400, 2401-2450, 2451-2500, 2501-2550, 2551-2600, 2601-2650, 2651-2700, 2701-2750, 2751-2800, 2801-2850, 2851-2900, 2901-2950, 2951-3000, 3001-3050, 3051-3100, 3101-3150, 3151-3200, 3201-3250, 3251-3300, 3301-3350, 3351-3400, 3401-3450, 3451-3500, 3501-3550, 3551-3600, 3601-3650, 3651-3700, 3701-3750, 3751-3800, 3801-3850, 3851-3900, 3901-3950, 3951-4000, 4001-4050, 4051-4100, 4101-4150, 4151-4200, 4201-4250, 4251-4300, 4301-4350, 4351-4400, 4401-4450, 4451-4500, 4501-4550, 4551-4600, 4601-4650, 4651-4700, 4701-4750, 4751-4800, 4801-4850, 4851-4900, 4901-4950, 4951-5000, 5001-5050, 5051-5100, 5101-5150, 5151-5200, 5201-5250, 5251-5300, 5301-5350, 5351-5400, 5401-5450, 5451-5500, 5501-5550, 5551-5600, 5601-5650, 5651-5700, 5701-5750, 5751-5800, 5801-5850, 5851-5900, 5901-5950, 5951-6000, 6001-6050, 6051-6100, 6101-6150, 6151-6200, 6201-6250, 6251-6300, 6301-6350, 6351-6400, 6401-6450, 6451-6500, 6501-6550, 6551-6600, 6601-6650, 6651-6700, 6701-6750, 6751-6800, 6801-6850, 6851-6900, 6901-6950, 6951-7000, 7001-7050, 7051-7100, 7101-7150, 7151-7200, 7201-7250, 7251-7300 or 7301 to the end of the cDNA sequence contained in ATCC Deposit No:Z, or the complementary strand thereto. In this context “about” includes the particularly recited range or a range larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. Preferably, these fragments encode a polypeptide which has a functional activity (e.g., biological activity). More preferably, these polynucleotides can be used as probes or primers as discussed herein. Polynucleotides which hybridize to one or more of these polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions are also encompassed by the invention, as are polypeptides encoded by these polynucleotides.

Moreover, representative examples of polynucleotide fragments of the invention comprise, or alternatively consist of, a nucleic acid sequence comprising one, two, three, four, five, six, seven, eight, nine, ten, or more of the above described polynucleotide fragments of the invention in combination with a polynucleotide sequence delineated in Table 1C column 6. Additional, representative examples of polynucleotide fragments of the invention comprise, or alternatively consist of, a nucleic acid sequence comprising one, two, three, four, five, six, seven, eight, nine, ten, or more of the above described polynucleotide fragments of the invention in combination with a polynucleotide sequence that is the complementary strand of a sequence delineated in column 6 of Table 1C. In further embodiments, the above-described polynucleotide fragments of the invention comprise, or alternatively consist of, sequences delineated in Table 1C, column 6, and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ID NO:B (see Table 1C, column 5). In additional embodiments, the above-described polynucleotide fragments of the invention comprise, or alternatively consist of, sequences delineated in Table 1C, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated Table 1C, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ID NO:A (see Table 1C, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more fragments of the sequences delineated in column 6 of Table 1C, and the polynucleotide sequence of SEQ ID NO:X (e.g., as defined in Table 1C, column 2) or fragments or variants thereof. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more fragments of the sequences delineated in column 6 of Table 1C which correspond to the same ATCC Deposit No:Z (see Table 1C, column 1), and the polynucleotide sequence of SEQ ID NO:X (e.g., as defined in Table 1A, 1B, or 1C) or fragments or variants thereof. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.

In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more fragments of the sequences delineated in the same row of column 6 of Table 1C, and the polynucleotide sequence of SEQ ID NO:X (e.g., as defined in Table 1A, 1B, or 1C) or fragments or variants thereof. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of the sequence of SEQ ID NO:X are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of a fragment or variant of the sequence of SEQ ID NO:X (e.g., as described herein) are directly contiguous Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3′ 10 polynucleotides of a fragment or variant of the sequence of SEQ ID NO:X and the 5′ 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table 1C are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C and the 5′ 10 polynucleotides of another sequence in column 6 are directly contiguous. In preferred embodiments, the 3′ 10 polynucleotides of one of the sequences delineated in column 6 of Table 1C is directly contiguous with the 5′ 10 polynucleotides of the next sequential exon delineated in Table 1C, column 6. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.

In the present invention, a “polypeptide fragment” refers to an amino acid sequence which is a portion of the amino acid sequence contained in SEQ ID NO:Y, is a portion of the mature form of SEQ ID NO:Y as defined in columns 14 and 15 of Table 1A, a portion of an amino acid sequence encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, is a portion of an amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:X, is a portion of an amino acid sequence encoded by the complement of the polynucleotide sequence in SEQ ID NO:X, is a portion of the amino acid sequence of a mature (secreted) polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or is a portion of an amino acid sequence encoded by the cDNA contained in ATCC Deposit No:Z. Protein (polypeptide) fragments may be “free-standing,” or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the invention, include, for example, fragments comprising, or alternatively consisting of, from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, 101-120, 121-140, 141-160, 161-180, 181-200, 201-220, 221-240, 241-260, 261-280, 281-300, 301-320, 321-340, 341-360, 361-380, 381-400, 401-420, 421-440, 441-460, 461-480, 481-500, 501-520, 521-540, 541-560, 561-580, 581-600, 601-620, 621-640, 641-660, 661-680, 681-700, 701-720, 721-740, 741-760, 761-780, 781-800, 801-820, 821-840, 841-860, 861-880, 881-900, 901-920, 921-940, 941-960, 961-980, 981-1000, 1001-1020, 1021-1040, 1041-1060, 1061-1080, 1081-1100, 1101-1120, 1121-1140, 1141-1160, 1161-1180, 1181-1200, 1201-1220, 1221-1240, 1241-1260, 1261-1280, 1281-1300, 1301-1320, 1321-1340, 1341-1360, 1361-1380, 1381-1400, 1401-1420, 1421-1440, or 1441 to the end of the coding region of cDNA and SEQ ID NO: Y. In a preferred embodiment, polypeptide fragments of the invention include, for example, fragments comprising, or alternatively consisting of, from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, 101-120, 121-140, 141-160, 161-180, 181-200, 201-220, 221-240, 241-260, 261-280, 281-300, 301-320, 321-340, 341-360, 361-380, 381-400, 401-420, 421-440, 441-460, 461-480, 481-500, 501-520, 521-540, 541-560, 561-580, 581-600, 601-620, 621-640, 641-660, 661-680, 681-700, 701-720, 721-740, 741-760, 761-780, 781-800, 801-820, 821-840, 841-860, 861-880, 881-900, 901-920, 921-940, 941-960, 961-980, 981-1000, 1001-1020, 1021-1040, 1041-1060, 1061-1080, 1081-1100, 1101-1120, 1121-1140, 1141-1160, 1161-1180, 1181-1200, 1201-1220, 1221-1240, 1241-1260, 1261-1280, 1281-1300, 1301-1320, 1321-1340, 1341-1360, 1361-1380, 1381-1400, 1401-1420, 1421-1440, or 1441 to the end of the coding region of SEQ ID NO:Y. Moreover, polypeptide fragments of the invention may be at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 100, 110, 120, 130, 140, or 150 amino acids in length. In this context “about” includes the particularly recited ranges or values, or ranges or values larger or smaller by several (5, 4, 3, 2, or 1) amino acids, at either extreme or at both extremes. Polynucleotides encoding these polypeptide fragments are also encompassed by the invention.

Even if deletion of one or more amino acids from the N-terminus of a protein results in modification of loss of one or more biological functions of the protein, other functional activities (e.g., biological activities, ability to multimerize, ability to bind a ligand) may still be retained. For example, the ability of shortened muteins to induce and/or bind to antibodies which recognize the complete or mature forms of the polypeptides generally will be retained when less than the majority of the residues of the complete or mature polypeptide are removed from the N-terminus. Whether a particular polypeptide lacking N-terminal residues of a complete polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art. It is not unlikely that a mutein with a large number of deleted N-terminal amino acid residues may retain some biological or immunogenic activities. In fact, peptides composed of as few as six amino acid residues may often evoke an immune response.

Accordingly, polypeptide fragments include the secreted protein as well as the mature form. Further preferred polypeptide fragments include the secreted protein or the mature form having a continuous series of deleted residues from the amino or the carboxy terminus, or both. For example, any number of amino acids, ranging from 1-60, can be deleted from the amino terminus of either the secreted polypeptide or the mature form. Similarly, any number of amino acids, ranging from 1-30, can be deleted from the carboxy terminus of the secreted protein or mature form. Furthermore, any combination of the above amino and carboxy terminus deletions are preferred. Similarly, polynucleotides encoding these polypeptide fragments are also preferred.

The present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of a polypeptide disclosed herein (e.g., a polypeptide of SEQ ID NO:Y, a polypeptide as defined in columns 14 and 15 of Table 1A, a polypeptide encoded by the polynucleotide sequence contained in SEQ ID NO:X or the complement thereof, a polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, a polypeptide encoded by the portion of SEQ ID NO:B as defined in column 6 of Table 1C, a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or a mature polypeptide encoded by the cDNA contained in ATCC Deposit No:Z). In particular, N-terminal deletions may be described by the general formula m-q, where q is a whole integer representing the total number of amino acid residues in a polypeptide of the invention (e.g., the polypeptide disclosed in SEQ ID NO:Y, the mature (secreted) portion of SEQ ID NO:Y as defined in columns 14 and 15 of Table 1A, or the polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2), and m is defined as any integer ranging from 2 to q-6. Polynucleotides encoding these polypeptides are also encompassed by the invention.

The present invention further provides polypeptides having one or more residues from the carboxy terminus of the amino acid sequence of a polypeptide disclosed herein (e.g., a polypeptide of SEQ ID NO:Y, the mature (secreted) portion of SEQ ID NO:Y as defined in columns 14 and 15 of Table 1A, a polypeptide encoded by the polynucleotide sequence contained in SEQ ID NO:X, a polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, a polypeptide encoded by the portion of SEQ ID NO:B as defined in column 6 of Table 1C, a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or a mature polypeptide encoded by the cDNA contained in ATCC Deposit No:Z). In particular, C-terminal deletions may be described by the general formula 1-n, where n is any whole integer ranging from 6 to q-1, and where n corresponds to the position of amino acid residue in a polypeptide of the invention. Polynucleotides encoding these polypeptides are also encompassed by the invention.

In addition, any of the above described N- or C-terminal deletions can be combined to produce a N- and C-terminal deleted polypeptide. The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini, which may be described generally as having residues m-n of a polypeptide encoded by SEQ ID NO:X (e.g., including, but not limited to, the preferred polypeptide disclosed as SEQ ID NO:Y, the mature (secreted) portion of SEQ ID NO:Y as defined in columns 14 and 15 of Table 1A, and the polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2), the cDNA contained in ATCC Deposit No:Z, and/or the complement thereof, where n and m are integers as described above. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Also as mentioned above, even if deletion of one or more amino acids from the C-terminus of a protein results in modification of loss of one or more biological functions of the protein, other functional activities (e.g., biological activities, ability to multimerize, ability to bind a ligand) may still be retained. For example the ability of the shortened mutein to induce and/or bind to antibodies which recognize the complete or mature forms of the polypeptide generally will be retained when less than the majority of the residues of the complete or mature polypeptide are removed from the C-terminus. Whether a particular polypeptide lacking C-terminal residues of a complete polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art. It is not unlikely that a mutein with a large number of deleted C-terminal amino acid residues may retain some biological or immunogenic activities. In fact, peptides composed of as few as six amino acid residues may often evoke an immune response.

The present application is also directed to proteins containing polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a polypeptide sequence set forth herein. In preferred embodiments, the application is directed to proteins containing polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to polypeptides having the amino acid sequence of the specific N- and C-terminal deletions. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Any polypeptide sequence encoded by, for example, the polynucleotide sequences set forth as SEQ ID NO:X or the complement thereof, (presented, for example, in Tables 1A and 2), the cDNA contained in ATCC Deposit No:Z, or the polynucleotide sequence as defined in column 6 of Table 1C, may be analyzed to determine certain preferred regions of the polypeptide. For example, the amino acid sequence of a polypeptide encoded by a polynucleotide sequence of SEQ ID NO:X (e.g., the polypeptide of SEQ ID NO:Y and the polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2) or the cDNA contained in ATCC Deposit No:Z may be analyzed using the default parameters of the DNASTAR computer algorithm (DNASTAR, Inc., 1228 S. Park St., Madison, Wis. 53715 USA; http://www.dnastar.com/).

Polypeptide regions that may be routinely obtained using the DNASTAR computer algorithm include, but are not limited to, Garnier-Robson alpha-regions, beta-regions, turn-regions, and coil-regions; Chou-Fasman alpha-regions, beta-regions, and turn-regions; Kyte-Doolittle hydrophilic regions and hydrophobic regions; Eisenberg alpha- and beta-amphipathic regions; Karplus-Schulz flexible regions; Emini surface-forming regions; and Jameson-Wolf regions of high antigenic index. Among highly preferred polynucleotides of the invention in this regard are those that encode polypeptides comprising regions that combine several structural features, such as several (e.g., 1, 2, 3 or 4) of the features set out above.

Additionally, Kyte-Doolittle hydrophilic regions and hydrophobic regions, Emini surface-forming regions, and Jameson-Wolf regions of high antigenic index (i.e., containing four or more contiguous amino acids having an antigenic index of greater than or equal to 1.5, as identified using the default parameters of the Jameson-Wolf program) can routinely be used to determine polypeptide regions that exhibit a high degree of potential for antigenicity. Regions of high antigenicity are determined from data by DNASTAR analysis by choosing values which represent regions of the polypeptide which are likely to be exposed on the surface of the polypeptide in an environment in which antigen recognition may occur in the process of initiation of an immune response.

Preferred polypeptide fragments of the invention are fragments comprising, or alternatively, consisting of, an amino acid sequence that displays a functional activity (e.g. biological activity) of the polypeptide sequence of which the amino acid sequence is a fragment. By a polypeptide displaying a “functional activity” is meant a polypeptide capable of one or more known functional activities associated with a full-length protein, such as, for example, biological activity, antigenicity, immunogenicity, and/or multimerization, as described herein.

Other preferred polypeptide fragments are biologically active fragments. Biologically active fragments are those exhibiting activity similar, but not necessarily identical, to an activity of the polypeptide of the present invention. The biological activity of the fragments may include an improved desired activity, or a decreased undesirable activity.

In preferred embodiments, polypeptides of the invention comprise, or alternatively consist of, one, two, three, four, five or more of the antigenic fragments of the polypeptide of SEQ ID NO:Y, or portions thereof. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Epitopes and Antibodies

The present invention encompasses polypeptides comprising, or alternatively consisting of, an epitope of: the polypeptide sequence shown in SEQ ID NO:Y; a polypeptide sequence encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide sequence encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2; the polypeptide sequence encoded by the portion of SEQ ID NO:B as defined in column 6 of Table 1C or the complement thereto; the polypeptide sequence encoded by the cDNA contained in ATCC Deposit No:Z; or the polypeptide sequence encoded by a polynucleotide that hybridizes to the sequence of SEQ ID NO:X, the complement of the sequence of SEQ ID NO:X, the complement of a portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, or the cDNA sequence contained in ATCC Deposit No:Z under stringent hybridization conditions or alternatively, under lower stringency hybridization as defined supra. The present invention further encompasses polynucleotide sequences encoding an epitope of a polypeptide sequence of the invention (such as, for example, the sequence disclosed in SEQ ID NO:X, or a fragment thereof), polynucleotide sequences of the complementary strand of a polynucleotide sequence encoding an epitope of the invention, and polynucleotide sequences which hybridize to the complementary strand under stringent hybridization conditions or alternatively, under lower stringency hybridization conditions defined supra.

The term “epitopes,” as used herein, refers to portions of a polypeptide having antigenic or immunogenic activity in an animal, preferably a mammal, and most preferably in a human. In a preferred embodiment, the present invention encompasses a polypeptide comprising an epitope, as well as the polynucleotide encoding this polypeptide. An “immunogenic epitope,” as used herein, is defined as a portion of a protein that elicits an antibody response in an animal, as determined by any method known in the art, for example, by the methods for generating antibodies described infra. (See, for example, Geysen et al., Proc. Natl. Acad. Sci. USA 81:3998-4002 (1983)). The term “antigenic epitope,” as used herein, is defined as a portion of a protein to which an antibody can immunospecifically bind its antigen as determined by any method well known in the art, for example, by the immunoassays described herein. Immunospecific binding excludes non-specific binding but does not necessarily exclude cross-reactivity with other antigens. Antigenic epitopes need not necessarily be immunogenic.

Fragments which function as epitopes may be produced by any conventional means. (See, e.g., Houghten, R. A., Proc. Natl. Acad. Sci. USA 82:5131-5135 (1985) further described in U.S. Pat. No. 4,631,211.)

In the present invention, antigenic epitopes preferably contain a sequence of at least 4, at least 5, at least 6, at least 7, more preferably at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, and, most preferably, between about 15 to about 30 amino acids. Preferred polypeptides comprising immunogenic or antigenic epitopes are at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acid residues in length. Additional non-exclusive preferred antigenic epitopes include the antigenic epitopes disclosed herein, as well as portions thereof. Antigenic epitopes are useful, for example, to raise antibodies, including monoclonal antibodies, that specifically bind the epitope. Preferred antigenic epitopes include the antigenic epitopes disclosed herein, as well as any combination of two, three, four, five or more of these antigenic epitopes. Antigenic epitopes can be used as the target molecules in immunoassays. (See, for instance, Wilson et al., Cell 37:767-778 (1984); Sutcliffe et al., Science 219:660-666 (1983)).

Non-limiting examples of epitopes of polypeptides that can be used to generate antibodies of the invention include a polypeptide comprising, or alternatively consisting of, at least one, two, three, four, five, six or more of the portion(s) of SEQ ID NO:Y specified in column 6 of Table 1B.1. These polypeptide fragments have been determined to bear antigenic epitopes of the proteins of the invention by the analysis of the Jameson-Wolf antigenic index which is included in the DNAStar suite of computer programs. By “comprise” it is intended that a polypeptide contains at least one, two, three, four, five, six or more of the portion(s) of SEQ ID NO:Y shown in column 6 of Table 1B.1, but it may contain additional flanking residues on either the amino or carboxyl termini of the recited portion. Such additional flanking sequences are preferably sequences naturally found adjacent to the portion; i.e., contiguous sequence shown in SEQ ID NO:Y. The flanking sequence may, however, be sequences from a heterolgous polypeptide, such as from another protein described herein or from a heterologous polypeptide not described herein. In particular embodiments, epitope portions of a polypeptide of the invention comprise one, two, three, or more of the portions of SEQ ID NO:Y shown in column 6 of Table 1B.1.

Similarly, immunogenic epitopes can be used, for example, to induce antibodies according to methods well known in the art. See, for instance, Sutcliffe et al., supra; Wilson et al., supra; Chow et al., Proc. Natl. Acad. Sci. USA 82:910-914; and Bittle et al., J. Gen. Virol. 66:2347-2354 (1985). Preferred immunogenic epitopes include the immunogenic epitopes disclosed herein, as well as any combination of two, three, four, five or more of these immunogenic epitopes. The polypeptides comprising one or more immunogenic epitopes may be presented for eliciting an antibody response together with a carrier protein, such as an albumin, to an animal system (such as rabbit or mouse), or, if the polypeptide is of sufficient length (at least about 25 amino acids), the polypeptide may be presented without a carrier. However, immunogenic epitopes comprising as few as 8 to 10 amino acids have been shown to be sufficient to raise antibodies capable of binding to, at the very least, linear epitopes in a denatured polypeptide (e.g., in Western blotting).

Epitope-bearing polypeptides of the present invention may be used to induce antibodies according to methods well known in the art including, but not limited to, in vivo immunization, in vitro immunization, and phage display methods. See, e.g., Sutcliffe et al., supra; Wilson et al., supra, and Bittle et al., J. Gen. Virol., 66:2347-2354 (1985). If in vivo immunization is used, animals may be immunized with free peptide; however, anti-peptide antibody titer may be boosted by coupling the peptide to a macromolecular carrier, such as keyhole limpet hemacyanin (KLH) or tetanus toxoid. For instance, peptides containing cysteine residues may be coupled to a carrier using a linker such as maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), while other peptides may be coupled to carriers using a more general linking agent such as glutaraldehyde. Animals such as rabbits, rats and mice are immunized with either free or carrier-coupled peptides, for instance, by intraperitoneal and/or intradermal injection of emulsions containing about 100 μg of peptide or carrier protein and Freund's adjuvant or any other adjuvant known for stimulating an immune response. Several booster injections may be needed, for instance, at intervals of about two weeks, to provide a useful titer of anti-peptide antibody which can be detected, for example, by ELISA assay using free peptide adsorbed to a solid surface. The titer of anti-peptide antibodies in serum from an immunized animal may be increased by selection of anti-peptide antibodies, for instance, by adsorption to the peptide on a solid support and elution of the selected antibodies according to methods well known in the art.

As one of skill in the art will appreciate, and as discussed above, the polypeptides of the present invention (e.g., those comprising an immunogenic or antigenic epitope) can be fused to heterologous polypeptide sequences. For example, polypeptides of the present invention (including fragments or variants thereof), may be fused with the constant domain of immunoglobulins (IgA, IgE, IgG, IgM), or portions thereof (CH1, CH2, CH3, or any combination thereof and portions thereof, resulting in chimeric polypeptides. By way of another non-limiting example, polypeptides and/or antibodies of the present invention (including fragments or variants thereof) may be fused with albumin (including but not limited to recombinant human serum albumin or fragments or variants thereof (see, e.g., U.S. Pat. No. 5,876,969, issued Mar. 2, 1999, EP Patent 0 413 622, and U.S. Pat. No. 5,766,883, issued Jun. 16, 1998, herein incorporated by reference in their entirety)). In a preferred embodiment, polypeptides and/or antibodies of the present invention (including fragments or variants thereof) are fused with the mature form of human serum albumin (i.e., amino acids 1-585 of human serum albumin as shown in FIGS. 1 and 2 of EP Patent 0 322 094) which is herein incorporated by reference in its entirety. In another preferred embodiment, polypeptides and/or antibodies of the present invention (including fragments or variants thereof) are fused with polypeptide fragments comprising, or alternatively consisting of, amino acid residues 1-z of human serum albumin, where z is an integer from 369 to 419, as described in U.S. Pat. No. 5,766,883 herein incorporated by reference in its entirety. Polypeptides and/or antibodies of the present invention (including fragments or variants thereof) may be fused to either the N- or C-terminal end of the heterologous protein (e.g., immunoglobulin Fc polypeptide or human serum albumin polypeptide). Polynucleotides encoding fusion proteins of the invention are also encompassed by the invention.

Such fusion proteins as those described above may facilitate purification and may increase half-life in vivo. This has been shown for chimeric proteins consisting of the first two domains of the human CD4-polypeptide and various domains of the constant regions of the heavy or light chains of mammalian immunoglobulins. See, e.g., EP 394,827; Traunecker et al., Nature, 331:84-86 (1988). Enhanced delivery of an antigen across the epithelial barrier to the immune system has been demonstrated for antigens (e.g., insulin) conjugated to an FcRn binding partner such as IgG or Fc fragments (see, e.g., PCT Publications WO 96/22024 and WO 99/04813). IgG fusion proteins that have a disulfide-linked dimeric structure due to the IgG portion desulfide bonds have also been found to be more efficient in binding and neutralizing other molecules than monomeric polypeptides or fragments thereof alone. See, e.g., Fountoulakis et al., J. Biochem., 270:3958-3964 (1995). Nucleic acids encoding the above epitopes can also be recombined with a gene of interest as an epitope tag (e.g., the hemagglutinin (HA) tag or flag tag) to aid in detection and purification of the expressed polypeptide. For example, a system described by Janknecht et al. allows for the ready purification of non-denatured fusion proteins expressed in human cell lines (Janknecht et al., 1991, Proc. Natl. Acad. Sci. USA 88:8972-897). In this system, the gene of interest is subcloned into a vaccinia recombination plasmid such that the open reading frame of the gene is translationally fused to an amino-terminal tag consisting of six histidine residues. The tag serves as a matrix binding domain for the fusion protein. Extracts from cells infected with the recombinant vaccinia virus are loaded onto Ni2+ nitriloacetic acid-agarose column and histidine-tagged proteins can be selectively eluted with imidazole-containing buffers.

Fusion Proteins

Any polypeptide of the present invention can he used to generate fusion proteins. For example, the polypeptide of the present invention, when fused to a second protein, can be used as an antigenic tag. Antibodies raised against the polypeptide of the present invention can be used to indirectly detect the second protein by binding to the polypeptide. Moreover, because secreted proteins target cellular locations based on trafficking signals, polypeptides of the present invention which are shown to be secreted can be used as targeting molecules once fused to other proteins.

Examples of domains that can be fused to polypeptides of the present invention include not only heterologous signal sequences, but also other heterologous functional regions. The fusion does not necessarily need to be direct, but may occur through linker sequences.

In certain preferred embodiments, proteins of the invention are fusion proteins comprising an amino acid sequence that is an N and/or C-terminal deletion of a polypeptide of the invention. In preferred embodiments, the invention is directed to a fusion protein comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98% or 99% identical to a polypeptide sequence of the invention. Polynucleotides encoding these proteins are also encompassed by the invention.

Moreover, fusion proteins may also be engineered to improve characteristics of the polypeptide of the present invention. For instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence during purification from the host cell or subsequent handling and storage. Also, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to facilitate handling of polypeptides are familiar and routine techniques in the art.

As one of skill in the art will appreciate that, as discussed above, polypeptides of the present invention, and epitope-bearing fragments thereof, can be combined with heterologous polypeptide sequences. For example, the polypeptides of the present invention may be fused with heterologous polypeptide sequences, for example, the polypeptides of the present invention may be fused with the constant domain of immunoglobulins (IgA, IgE, IgG, IgM) or portions thereof (CH1, CH2, CH3, and any combination thereof, including both entire domains and portions thereof), or albumin (including, but not limited to, native or recombinant human albumin or fragments or variants thereof (see, e.g., U.S. Pat. No. 5,876,969, issued Mar. 2, 1999, EP Patent 0 413 622, and U.S. Pat. No. 5,766,883, issued Jun. 16, 1998, herein incorporated by reference in their entirety)), resulting in chimeric polypeptides. For example, EP-A-O 464 533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobulin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is beneficial in therapy and diagnosis, and thus can result in, for example, improved pharmacokinetic properties (EP-A 0232 262). Alternatively, deleting the Fc part after the fusion protein has been expressed, detected, and purified, would be desired. For example, the Fc portion may hinder therapy and diagnosis if the fusion protein is used as an antigen for immunizations. In drug discovery, for example, human proteins, such as hIL-5, have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. See, D. Bennett et al., J. Molecular Recognition 8:52-58 (1995); K. Johanson et al., J. Biol. Chem. 270:9459-9471 (1995).

Moreover, the polypeptides of the present invention can be fused to marker sequences, such as a polypeptide which facilitates purification of the fused polypeptide. In preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, Calif., 91311), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. Another peptide tag useful for purification, the “HA” tag, corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., Cell 37:767 (1984)).

Additional fusion proteins of the invention may be generated through the techniques of gene-shuffling, motif-shuffling, exon-shuffling, and/or codon-shuffling (collectively referred to as “DNA shuffling”). DNA shuffling may be employed to modulate the activities of polypeptides of the invention, such methods can be used to generate polypeptides with altered activity, as well as agonists and antagonists of the polypeptides. See, generally, U.S. Pat. Nos. 5,605,793; 5,811,238; 5,830,721; 5,834,252; and 5,837,458, and Patten et al., Curr. Opinion Biotechnol. 8:724-33 (1997); Harayama, Trends Biotechnol. 16(2):76-82 (1998); Hansson, et al., J. Mol. Biol. 287:265-76 (1999); and Lorenzo and Blasco, Biotechniques 24(2):308-13 (1998) (each of these patents and publications are hereby incorporated by reference in its entirety). In one embodiment, alteration of polynucleotides corresponding to SEQ ID NO:X and the polypeptides encoded by these polynucleotides may be achieved by DNA shuffling. DNA shuffling involves the assembly of two or more DNA segments by homologous or site-specific recombination to generate variation in the polynucleotide sequence. In another embodiment, polynucleotides of the invention, or the encoded polypeptides, may be altered by being subjected to random mutagenesis by error-prone PCR, random nucleotide insertion or other methods prior to recombination. In another embodiment, one or more components, motifs, sections, parts, domains, fragments, etc., of a polynucleotide encoding a polypeptide of the invention may be recombined with one or more components, motifs, sections, parts, domains, fragments, etc. of one or more heterologous molecules.

Thus, any of these above fusions can be engineered using the polynucleotides or the polypeptides of the present invention.

Recombinant and Synthetic Production of Polypeptides of the Invention

The present invention also relates to vectors containing the polynucleotide of the present invention, host cells, and the production of polypeptides by synthetic and recombinant techniques. The vector may be, for example, a phage, plasmid, viral, or retroviral vector. Retroviral vectors may be replication competent or replication defective. In the latter case, viral propagation generally will occur only in complementing host cells.

The polynucleotides of the invention may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid. If the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.

The polynucleotide insert should be operatively linked to an appropriate promoter, such as the phage lambda PL promoter, the E. coli lac, trp, phoA and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters will be known to the skilled artisan. The expression constructs will further contain sites for transcription initiation, termination, and, in the transcribed region, a ribosome binding site for translation. The coding portion of the transcripts expressed by the constructs will preferably include a translation initiating codon at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.

As indicated, the expression vectors will preferably include at least one selectable marker. Such markers include dihydrofolate reductase, G418, glutamine synthase, or neomycin resistance for eukaryotic cell culture, and tetracycline, kanamycin or ampicillin resistance genes for culturing in E. coli and other bacteria. Representative examples of appropriate hosts include, but are not limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells (e.g., Saccharomyces cerevisiae or Pichia pastoris (ATCC Accession No. 201178)); insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS, 293, and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.

Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from QIAGEN, Inc.; pBluescript vectors, Phagescript vectors, pNH8A, pNH16a, pNH18A, pNH46A, available from Stratagene Cloning Systems, Inc.; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 available from Pharmacia Biotech, Inc. Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Preferred expression vectors for use in yeast systems include, but are not limited to pYES2, pYD1, pTEF1/Zeo, pYES2/GS, pPICZ, pGAPZ, pGAPZalph, pPIC9, pPIC3.5, pHIL-D2, pHIL-S1, pPIC3.5K, pPIC9K, and PAO815 (all available from Invitrogen, Carlbad, Calif.). Other suitable vectors will be readily apparent to the skilled artisan.

Vectors which use glutamine synthase (GS) or DHFR as the selectable markers can be amplified in the presence of the drugs methionine sulphoximine or methotrexate, respectively. An advantage of glutamine synthase based vectors are the availability of cell lines (e.g., the murine myeloma cell line, NS0) which are glutamine synthase negative. Glutamine synthase expression systems can also function in glutamine synthase expressing cells (e.g., Chinese Hamster Ovary (CHO) cells) by providing additional inhibitor to prevent the functioning of the endogenous gene. A glutamine synthase expression system and components thereof are detailed in PCT publications: WO87/04462; WO86/05807; WO89/01036; WO89/10404; and WO91/06657, which are hereby incorporated in their entireties by reference herein. Additionally, glutamine synthase expression vectors can be obtained from Lonza Biologics, Inc. (Portsmouth, N.H.). Expression and production of monoclonal antibodies using a GS expression system in murine myeloma cells is described in Bebbington et al., Bio/technology 10:169(1992) and in Biblia and Robinson Biotechnol. Prog. 11:1 (1995) which are herein incorporated by reference.

The present invention also relates to host cells containing the above-described vector constructs described herein, and additionally encompasses host cells containing nucleotide sequences of the invention that are operably associated with one or more heterologous control regions (e.g., promoter and/or enhancer) using techniques known of in the art. The host cell can be a higher eukaryotic cell, such as a mammalian cell (e.g., a human derived cell), or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. A host strain may be chosen which modulates the expression of the inserted gene sequences, or modifies and processes the gene product in the specific fashion desired. Expression from certain promoters can be elevated in the presence of certain inducers; thus expression of the genetically engineered polypeptide may be controlled. Furthermore, different host cells have characteristics and specific mechanisms for the translational and post-translational processing and modification (e.g., phosphorylation, cleavage) of proteins. Appropriate cell lines can be chosen to ensure the desired modifications and processing of the foreign protein expressed.

Introduction of the nucleic acids and nucleic acid constructs of the invention into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, or other methods. Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology (1986). It is specifically contemplated that the polypeptides of the present invention may in fact be expressed by a host cell lacking a recombinant vector.

In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., the coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous polynucleotide sequences via homologous recombination (see, e.g., U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication Number WO 96/29411; International Publication Number WO 94/12650; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).

Polypeptides of the invention can be recovered and purified from recombinant cell cultures by well-known methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography (“HPLC”) is employed for purification.

Polypeptides of the present invention can also be recovered from: products purified from natural sources, including bodily fluids, tissues and cells, whether directly isolated or cultured; products of chemical synthetic procedures; and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect, and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non-glycosylated. In addition, polypeptides of the invention may also include an initial modified methionine residue, in some cases as a result of host-mediated processes. Thus, it is well known in the art that the N-terminal methionine encoded by the translation initiation codon generally is removed with high efficiency from any protein after translation in all eukaryotic cells. While the N-terminal methionine on most proteins also is efficiently removed in most prokaryotes, for some proteins, this prokaryotic removal process is inefficient, depending on the nature of the amino acid to which the N-terminal methionine is covalently linked.

In one embodiment, the yeast Pichia pastoris is used to express polypeptides of the invention in a eukaryotic system. Pichia pastoris is a methylotrophic yeast which can metabolize methanol as its sole carbon source. A main step in the methanol metabolization pathway is the oxidation of methanol to formaldehyde using O₂. This reaction is catalyzed by the enzyme alcohol oxidase. In order to metabolize methanol as its sole carbon source, Pichia pastoris must generate high levels of alcohol oxidase due, in part, to the relatively low affinity of alcohol oxidase for O₂. Consequently, in a growth medium depending on methanol as a main carbon source, the promoter region of one of the two alcohol oxidase genes (AOX1) is highly active. In the presence of methanol, alcohol oxidase produced from the AOX1 gene comprises up to approximately 30% of the total soluble protein in Pichia pastoris. See Ellis, S. B., et al., Mol. Cell. Biol. 5:1111-21 (1985); Koutz, P. J, et al., Yeast 5:167-77 (1989); Tschopp, J. F., et al., Nucl. Acids Res. 15:3859-76 (1987). Thus, a heterologous coding sequence, such as, for example, a polynucleotide of the present invention, under the transcriptional regulation of all or part of the AOX1 regulatory sequence is expressed at exceptionally high levels in Pichia yeast grown in the presence of methanol.

In one example, the plasmid vector pPIC9K is used to express DNA encoding a polypeptide of the invention, as set forth herein, in a Pichea yeast system essentially as described in “Pichia Protocols: Methods in Molecular Biology,” D. R. Higgins and J. Cregg, eds. The Humana Press, Totowa, N.J., 1998. This expression vector allows expression and secretion of a polypeptide of the invention by virtue of the strong AOX1 promoter linked to the Pichia pastoris alkaline phosphatase (PHO) secretory signal peptide (i.e., leader) located upstream of a multiple cloning site.

Many other yeast vectors could be used in place of pPIC9K, such as, pYES2, pYD1, pTEF1/Zeo, pYES2/GS, pPICZ, pGAPZ, pGAPZalpha, pPIC9, pPIC3.5, pHIL-D2, pHIL-S1, pPIC3.5K, and PAO815, as one skilled in the art would readily appreciate, as long as the proposed expression construct provides appropriately located signals for transcription, translation, secretion (if desired), and the like, including an in-frame AUG as required.

In another embodiment, high-level expression of a heterologous coding sequence, such as, for example, a polynucleotide of the present invention, may be achieved by cloning the heterologous polynucleotide of the invention into an expression vector such as, for example, pGAPZ or pGAPZalpha, and growing the yeast culture in the absence of methanol.

In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous polynucleotide sequences via homologous recombination (see, e.g., U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication No. WO 96/29411, published Sep. 26, 1996; International Publication No. WO 94/12650, published Aug. 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).

In addition, polypeptides of the invention can be chemically synthesized using techniques known in the art (e.g., see Creighton, 1983, Proteins: Structures and Molecular Principles, W.H. Freeman & Co., N.Y., and Hunkapiller et al., Nature, 310:105-111 (1984)). For example, a polypeptide corresponding to a fragment of a polypeptide can be synthesized by use of a peptide synthesizer. Furthermore, if desired, nonclassical amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the polypeptide sequence. Non-classical amino acids include, but are not limited to, to the D-isomers of the common amino acids, 2,4-diaminobutyric acid, a-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, g-Abu, e-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, b-alanine, fluoro-amino acids, designer amino acids such as b-methyl amino acids, Ca-methyl amino acids, Na-methyl amino acids, and amino acid analogs in general. Furthermore, the amino acid can be D (dextrorotary) or L (levorotary).

The invention encompasses polypeptides of the present invention which are differentially modified during or after translation, e.g., by glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to an antibody molecule or other cellular ligand, etc. Any of numerous chemical modifications may be carried out by known techniques, including but not limited, to specific chemical cleavage by cyanogen bromide, trypsin, chymotrypsin, papain, V8 protease, NaBH₄; acetylation, formylation, oxidation, reduction; metabolic synthesis in the presence of tunicamycin; etc.

Additional post-translational modifications encompassed by the invention include, for example, e.g., N-linked or O-linked carbohydrate chains, processing of N-terminal or C-terminal ends), attachment of chemical moieties to the amino acid backbone, chemical modifications of N-linked or O-linked carbohydrate chains, and addition or deletion of an N-terminal methionine residue as a result of procaryotic host cell expression. The polypeptides may also be modified with a detectable label, such as an enzymatic, fluorescent, isotopic or affinity label to allow for detection and isolation of the protein.

Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin; and examples of suitable radioactive material include iodine (¹²¹I, ¹²¹I, ¹²⁵I, ¹³¹I), carbon (¹⁴C), sulfur (³⁵S), tritium (³H), indium (¹¹¹In, ¹¹²In, ^(113m)In, ^(115m)In), technetium (⁹⁹Tc, ^(99m)Tc), thallium (²⁰¹Ti), gallium (⁶⁸Ga, ⁶⁷Ga), palladium (¹⁰³Pd), molybdenum (⁹⁹Mo), xenon (¹³³Xe), fluorine (¹⁸F), ¹⁵³Sm, ¹⁷⁷Lu, ¹⁵⁹Gd, ¹⁴⁹Pm, ¹⁴⁰La, ¹⁷⁵Yb, ¹⁶⁶Ho, ⁹⁰Y, ⁴⁷Sc, ¹⁸⁶Re, ⁸⁸Re, ¹⁴²Pr, ¹⁰⁵Rh, and ⁹⁷Ru.

In specific embodiments, a polypeptide of the present invention or fragment or variant thereof is attached to macrocyclic chelators that associate with radiometal ions, including but not limited to, ¹⁷⁷Lu, ⁹⁰Y, ¹⁶⁶Ho, and ¹⁵³Sm, to polypeptides. In a preferred embodiment, the radiometal ion associated with the macrocyclic chelators is ¹¹¹In. In another preferred embodiment, the radiometal ion associated with the macrocyclic chelator is ⁹⁰Y. In specific embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N′,N″,N′″-tetraacetic acid (DOTA). In other specific embodiments, DOTA is attached to an antibody of the invention or fragment thereof via a linker molecule. Examples of linker molecules useful for conjugating DOTA to a polypeptide are commonly known in the art—see, for example, DeNardo et al., Clin Cancer Res. 4(10):2483-90 (1998); Peterson et al., Bioconjug. Chem. 10(4):553-7 (1999); and Zimmerman et al, Nucl. Med. Biol. 26(8):943-50 (1999); which are hereby incorporated by reference in their entirety.

As mentioned, the proteins of the invention may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Polypeptides of the invention may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. (See, for instance, PROTEINS—STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993); POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, pgs. 1-12 (1983); Seifter et al., Meth. Enzymol. 182:626-646 (1990); Rattan et al., Ann. N.Y. Acad. Sci. 663:48-62 (1992)).

Also provided by the invention are chemically modified derivatives of the polypeptides of the invention which may provide additional advantages such as increased solubility, stability and circulating time of the polypeptide, or decreased immunogenicity (see U.S. Pat. No. 4,179,337). The chemical moieties for derivitization may be selected from water soluble polymers such as polyethylene glycol, ethylene glycol/propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol and the like. The polypeptides may be modified at random positions within the molecule, or at predetermined positions within the molecule and may include one, two, three or more attached chemical moieties.

The polymer may be of any molecular weight, and may be branched or unbranched. For polyethylene glycol, the preferred molecular weight is between about 1 kDa and about 100 kDa (the term “about” indicating that in preparations of polyethylene glycol, some molecules will weigh more, some less, than the stated molecular weight) for ease in handling and manufacturing. Other sizes may be used, depending on the desired therapeutic profile (e.g., the duration of sustained release desired, the effects, if any on biological activity, the ease in handling, the degree or lack of antigenicity and other known effects of the polyethylene glycol to a therapeutic protein or analog). For example, the polyethylene glycol may have an average molecular weight of about 200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, 20,000, 25,000, 30,000, 35,000, 40,000, 45,000, 50,000, 55,000, 60,000, 65,000, 70,000, 75,000, 80,000, 85,000, 90,000, 95,000, or 100,000 kDa.

As noted above, the polyethylene glycol may have a branched structure. Branched polyethylene glycols are described, for example, in U.S. Pat. No. 5,643,575; Morpurgo et al., Appl. Biochem. Biotechnol. 56:59-72 (1996); Vorobjev et al., Nucleosides Nucleotides 18:2745-2750 (1999); and Caliceti et al., Bioconjug. Chem. 10:638-646 (1999), the disclosures of each of which are incorporated herein by reference.

The polyethylene glycol molecules (or other chemical moieties) should be attached to the protein with consideration of effects on functional or antigenic domains of the protein. There are a number of attachment methods available to those skilled in the art, such as, for example, the method disclosed in EP 0 401 384 (coupling PEG to G-CSF), herein incorporated by reference; see also Malik et al., Exp. Hematol. 20:1028-1035 (1992), reporting pegylation of GM-CSF using tresyl chloride. For example, polyethylene glycol may be covalently bound through amino acid residues via a reactive group, such as a free amino or carboxyl group. Reactive groups are those to which an activated polyethylene glycol molecule may be bound. The amino acid residues having a free amino group may include lysine residues and the N-terminal amino acid residues; those having a free carboxyl group may include aspartic acid residues glutamic acid residues and the C-terminal amino acid residue. Sulfhydryl groups may also be used as a reactive group for attaching the polyethylene glycol molecules. Preferred for therapeutic purposes is attachment at an amino group, such as attachment at the N-terminus or lysine group.

As suggested above, polyethylene glycol may be attached to proteins via linkage to any of a number of amino acid residues. For example, polyethylene glycol can be linked to proteins via covalent bonds to lysine, histidine, aspartic acid, glutamic acid, or cysteine residues. One or more reaction chemistries may be employed to attach polyethylene glycol to specific amino acid residues (e.g., lysine, histidine, aspartic acid, glutamic acid, or cysteine) of the protein or to more than one type of amino acid residue (e.g., lysine, histidine, aspartic acid, glutamic acid, cysteine and combinations thereof) of the protein.

One may specifically desire proteins chemically modified at the N-terminus. Using polyethylene glycol as an illustration of the present composition, one may select from a variety of polyethylene glycol molecules (by molecular weight, branching, etc.), the proportion of polyethylene glycol molecules to protein (polypeptide) molecules in the reaction mix, the type of pegylation reaction to be performed, and the method of obtaining the selected N-terminally pegylated protein. The method of obtaining the N-terminally pegylated preparation (i.e., separating this moiety from other monopegylated moieties if necessary) may be by purification of the N-terminally pegylated material from a population of pegylated protein molecules. Selective proteins chemically modified at the N-terminus modification may be accomplished by reductive alkylation which exploits differential reactivity of different types of primary amino groups (lysine versus the N-terminal) available for derivatization in a particular protein. Under the appropriate reaction conditions, substantially selective derivatization of the protein at the N-terminus with a carbonyl group containing polymer is achieved.

As indicated above, pegylation of the proteins of the invention may be accomplished by any number of means. For example, polyethylene glycol may be attached to the protein either directly or by an intervening linker. Linkerless systems for attaching polyethylene glycol to proteins are described in Delgado et al., Crit. Rev. Thera. Drug Carrier Sys. 9:249-304 (1992); Francis et al., Intern. J. of Hematol. 68:1-18 (1998); U.S. Pat. No. 4,002,531; U.S. Pat. No. 5,349,052; WO 95/06058; and WO 98/32466, the disclosures of each of which are incorporated herein by reference.

One system for attaching polyethylene glycol directly to amino acid residues of proteins without an intervening linker employs tresylated MPEG, which is produced by the modification of monmethoxy polyethylene glycol (MPEG) using tresylchloride (ClSO₂CH₂CF₃). Upon reaction of protein with tresylated MPEG, polyethylene glycol is directly attached to amine groups of the protein. Thus, the invention includes protein-polyethylene glycol conjugates produced by reacting proteins of the invention with a polyethylene glycol molecule having a 2,2,2-trifluoreothane sulphonyl group.

Polyethylene glycol can also be attached to proteins using a number of different intervening linkers. For example, U.S. Pat. No. 5,612,460, the entire disclosure of which is incorporated herein by reference, discloses urethane linkers for connecting polyethylene glycol to proteins. Protein-polyethylene glycol conjugates wherein the polyethylene glycol is attached to the protein by a linker can also be produced by reaction of proteins with compounds such as MPEG-succinimidylsuccinate, MPEG activated with 1,1′-carbonyldiimidazole, MPEG-2,4,5-trichloropenylcarbonate, MPEG-p-nitrophenolcarbonate, and various MPEG-succinate derivatives. A number of additional polyethylene glycol derivatives and reaction chemistries for attaching polyethylene glycol to proteins are described in International Publication No. WO 98/32466, the entire disclosure of which is incorporated herein by reference. Pegylated protein products produced using the reaction chemistries set out herein are included within the scope of the invention.

The number of polyethylene glycol moieties attached to each protein of the invention (i.e., the degree of substitution) may also vary. For example, the pegylated proteins of the invention may be linked, on average, to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 17, 20, or more polyethylene glycol molecules. Similarly, the average degree of substitution within ranges such as 1-3, 2-4, 3-5, 4-6, 5-7, 6-8, 7-9, 8-10, 9-11, 10-12, 11-13, 12-14, 13-15, 14-16, 15-17, 16-18, 17-19, or 18-20 polyethylene glycol moieties per protein molecule. Methods for determining the degree of substitution are discussed, for example, in Delgado et al., Crit. Rev. Thera. Drug Carrier Sys. 9:249-304 (1992).

The polypeptides of the invention can be recovered and purified from chemical synthesis and recombinant cell cultures by standard methods which include, but are not limited to, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography (“HPLC”) is employed for purification. Well known techniques for refolding protein may be employed to regenerate active conformation when the polypeptide is denatured during isolation and/or purification.

The polypeptides of the invention may be in monomers or multimers (i.e., dimers, trimers, tetramers and higher multimers). Accordingly, the present invention relates to monomers and multimers of the polypeptides of the invention, their preparation, and compositions (preferably, Therapeutics) containing them. In specific embodiments, the polypeptides of the invention are monomers, dimers, trimers or tetramers. In additional embodiments, the multimers of the invention are at least dimers, at least trimers, or at least tetramers.

Multimers encompassed by the invention may be homomers or heteromers. As used herein, the term homomer refers to a multimer containing only polypeptides corresponding to a protein of the invention (e.g., the amino acid sequence of SEQ ID NO:Y, an amino acid sequence encoded by SEQ ID NO:X or the complement of SEQ ID NO:X, the amino acid sequence encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, and/or an amino acid sequence encoded by cDNA contained in ATCC Deposit No:Z (including fragments, variants, splice variants, and fusion proteins, corresponding to these as described herein)). These homomers may contain polypeptides having identical or different amino acid sequences. In a specific embodiment, a homomer of the invention is a multimer containing only polypeptides having an identical amino acid sequence. In another specific embodiment, a homomer of the invention is a multimer containing polypeptides having different amino acid sequences. In specific embodiments, the multimer of the invention is a homodimer (e.g., containing two polypeptides having identical or different amino acid sequences) or a homotrimer (e.g., containing three polypeptides having identical and/or different amino acid sequences). In additional embodiments, the homomeric multimer of the invention is at least a homodimer, at least a homotrimer, or at least a homotetramer.

As used herein, the term heteromer refers to a multimer containing one or more heterologous polypeptides (i.e., polypeptides of different proteins) in addition to the polypeptides of the invention. In a specific embodiment, the multimer of the invention is a heterodimer, a heterotrimer, or a heterotetramer. In additional embodiments, the heteromeric multimer of the invention is at least a heterodimer, at least a heterotrimer, or at least a heterotetramer.

Multimers of the invention may be the result of hydrophobic, hydrophilic, ionic and/or covalent associations and/or may be indirectly linked by, for example, liposome formation. Thus, in one embodiment, multimers of the invention, such as, for example, homodimers or homotrimers, are formed when polypeptides of the invention contact one another in solution. In another embodiment, heteromultimers of the invention, such as, for example, heterotrimers or heterotetramers, are formed when polypeptides of the invention contact antibodies to the polypeptides of the invention (including antibodies to the heterologous polypeptide sequence in a fusion protein of the invention) in solution. In other embodiments, multimers of the invention are formed by covalent associations with and/or between the polypeptides of the invention. Such covalent associations may involve one or more amino acid residues contained in the polypeptide sequence (e.g., that recited in SEQ ID NO:Y, encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, and/or encoded by the cDNA contained in ATCC Deposit No:Z). In one instance, the covalent associations are cross-linking between cysteine residues located within the polypeptide sequences which interact in the native (i.e., naturally occurring) polypeptide. In another instance, the covalent associations are the consequence of chemical or recombinant manipulation. Alternatively, such covalent associations may involve one or more amino acid residues contained in the heterologous polypeptide sequence in a fusion protein. In one example, covalent associations are between the heterologous sequence contained in a fusion protein of the invention (see, e.g., U.S. Pat. No. 5,478,925). In a specific example, the covalent associations are between the heterologous sequence contained in a Fc fusion protein of the invention (as described herein). In another specific example, covalent associations of fusion proteins of the invention are between heterologous polypeptide sequence from another protein that is capable of forming covalently associated multimers, such as for example, osteoprotegerin (see, e.g., International Publication NO: WO 98/49305, the contents of which are herein incorporated by reference in its entirety). In another embodiment, two or more polypeptides of the invention are joined through peptide linkers. Examples include those peptide linkers described in U.S. Pat. No. 5,073,627 (hereby incorporated by reference). Proteins comprising multiple polypeptides of the invention separated by peptide linkers may be produced using conventional recombinant DNA technology.

Another method for preparing multimer polypeptides of the invention involves use of polypeptides of the invention fused to a leucine zipper or isoleucine zipper polypeptide sequence. Leucine zipper and isoleucine zipper domains are polypeptides that promote multimerization of the proteins in which they are found. Leucine zippers were originally identified in several DNA-binding proteins (Landschulz et al., Science 240:1759, (1988)), and have since been found in a variety of different proteins. Among the known leucine zippers are naturally occurring peptides and derivatives thereof that dimerize or trimerize. Examples of leucine zipper domains suitable for producing soluble multimeric proteins of the invention are those described in PCT application WO 94/10308, hereby incorporated by reference. Recombinant fusion proteins comprising a polypeptide of the invention fused to a polypeptide sequence that dimerizes or trimerizes in solution are expressed in suitable host cells, and the resulting soluble multimeric fusion protein is recovered from the culture supernatant using techniques known in the art.

Trimeric polypeptides of the invention may offer the advantage of enhanced biological activity. Preferred leucine zipper moieties and isoleucine moieties are those that preferentially form trimers. One example is a leucine zipper derived from lung surfactant protein D (SPD), as described in Hoppe et al. (FEBS Letters 344:191, (1994)) and in U.S. patent application Ser. No. 08/446,922, hereby incorporated by reference. Other peptides derived from naturally occurring trimeric proteins may be employed in preparing trimeric polypeptides of the invention.

In another example, proteins of the invention are associated by interactions between Flag® polypeptide sequence contained in fusion proteins of the invention containing Flag® polypeptide sequence. In a further embodiment, proteins of the invention are associated by interactions between heterologous polypeptide sequence contained in Flag® fusion proteins of the invention and anti-Flag® antibody.

The multimers of the invention may be generated using chemical techniques known in the art. For example, polypeptides desired to be contained in the multimers of the invention may be chemically cross-linked using linker molecules and linker molecule length optimization techniques known in the art (see, e.g., U.S. Pat. No. 5,478,925, which is herein incorporated by reference in its entirety). Additionally, multimers of the invention may be generated using techniques known in the art to form one or more inter-molecule cross-links between the cysteine residues located within the sequence of the polypeptides desired to be contained in the multimer (see, e.g., U.S. Pat. No. 5,478,925, which is herein incorporated by reference in its entirety). Further, polypeptides of the invention may be routinely modified by the addition of cysteine or biotin to the C-terminus or N-terminus of the polypeptide and techniques known in the art may be applied to generate multimers containing one or more of these modified polypeptides (see, e.g., U.S. Pat. No. 5,478,925, which is herein incorporated by reference in its entirety). Additionally, techniques known in the art may be applied to generate liposomes containing the polypeptide components desired to be contained in the multimer of the invention (see, e.g., U.S. Pat. No. 5,478,925, which is herein incorporated by reference in its entirety).

Alternatively, multimers of the invention may be generated using genetic engineering techniques known in the art. In one embodiment, polypeptides contained in multimers of the invention are produced recombinantly using fusion protein technology described herein or otherwise known in the art (see, e.g., U.S. Pat. No. 5,478,925, which is herein incorporated by reference in its entirety). In a specific embodiment, polynucleotides coding for a homodimer of the invention are generated by ligating a polynucleotide sequence encoding a polypeptide of the invention to a sequence encoding a linker polypeptide and then further to a synthetic polynucleotide encoding the translated product of the polypeptide in the reverse orientation from the original C-terminus to the N-terminus (lacking the leader sequence) (see, e.g., U.S. Pat. No. 5,478,925, which is herein incorporated by reference in its entirety). In another embodiment, recombinant techniques described herein or otherwise known in the art are applied to generate recombinant polypeptides of the invention which contain a transmembrane domain (or hydrophobic or signal peptide) and which can be incorporated by membrane reconstitution techniques into liposomes (see, e.g., U.S. Pat. No. 5,478,925, which is herein incorporated by reference in its entirety).

Antibodies

Further polypeptides of the invention relate to antibodies and T-cell antigen receptors (TCR) which immunospecifically bind a polypeptide, polypeptide fragment, or variant of the invention (e.g., a polypeptide or fragment or variant of the amino acid sequence of SEQ ID NO:Y or a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or an epitope, of the present invention) as determined by immunoassays well known in the art for assaying specific antibody-antigen binding. Antibodies of the invention include, but are not limited to, polyclonal, monoclonal, multispecific, human, humanized or chimeric antibodies, single chain antibodies, Fab fragments, F(ab′,) fragments, fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies (including, e.g., anti-Id antibodies to antibodies of the invention), intracellularly-made antibodies (i.e., intrabodies), and epitope-binding fragments of any of the above. The term “antibody,” as used herein, refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds an antigen. The immunoglobulin molecules of the invention can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass of immunoglobulin molecule. In preferred embodiments, the immunoglobulin molecules of the invention are IgG1. In other preferred embodiments, the immunoglobulin molecules of the invention are IgG4.

Most preferably the antibodies are human antigen-binding antibody fragments of the present invention and include, but are not limited to, Fab, Fab′ and F(ab′)2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv) and fragments comprising either a VL or VH domain. Antigen-binding antibody fragments, including single-chain antibodies, may comprise the variable region(s) alone or in combination with the entirety or a portion of the following: hinge region, CH1, CH2, and CH3 domains. Also included in the invention are antigen-binding fragments also comprising any combination of variable region(s) with a hinge region, CH1, CH2, and CH3 domains. The antibodies of the invention may be from any animal origin including birds and mammals. Preferably, the antibodies are human, murine (e.g., mouse and rat), donkey, ship rabbit, goat, guinea pig, camel, horse, or chicken. As used herein, “human” antibodies include antibodies having the amino acid sequence of a human immunoglobulin and include antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulin and that do not express endogenous immunoglobulins, as described infra and, for example in, U.S. Pat. No. 5,939,598 by Kucherlapati et al.

The antibodies of the present invention may be monospecific, bispecific, trispecific or of greater multispecificity. Multispecific antibodies may be specific for different epitopes of a polypeptide of the present invention or may be specific for both a polypeptide of the present invention as well as for a heterologous epitope, such as a heterologous polypeptide or solid support material. See, e.g., PCT publications WO 93/17715; WO 92/08802; WO 91/00360; WO 92/05793; Tutt, et al., J. Immunol. 147:60-69 (1991); U.S. Pat. Nos. 4,474,893; 4,714,681; 4,925,648; 5,573,920; 5,601,819; Kostelny et al., J. Immunol. 148:1547-1553 (1992).

Antibodies of the present invention may be described or specified in terms of the epitope(s) or portion(s) of a polypeptide of the present invention which they recognize or specifically bind. The epitope(s) or polypeptide portion(s) may be specified as described herein, e.g., by N-terminal and C-terminal positions, or by size in contiguous amino acid residues, or listed in the Tables and Figures. Preferred epitopes of the invention include the predicted epitopes shown in column 6 of Table 1B.1, as well as polynucleotides that encode these epitopes. Antibodies which specifically bind any epitope or polypeptide of the present invention may also be excluded. Therefore, the present invention includes antibodies that specifically bind polypeptides of the present invention, and allows for the exclusion of the same.

Antibodies of the present invention may also be described or specified in terms of their cross-reactivity. Antibodies that do not bind any other analog, ortholog, or homolog of a polypeptide of the present invention are included. Antibodies that bind polypeptides with at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55%, and at least 50% identity (as calculated using methods known in the art and described herein) to a polypeptide of the present invention are also included in the present invention. In specific embodiments, antibodies of the present invention cross-react with murine, rat and/or rabbit homologs of human proteins and the corresponding epitopes thereof. Antibodies that do not bind polypeptides with less than 95%, less than 90%, less than 85%, less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 55%, and less than 50% identity (as calculated using methods known in the art and described herein) to a polypeptide of the present invention are also included in the present invention. In a specific embodiment, the above-described cross-reactivity is with respect to any single specific antigenic or immunogenic polypeptide, or combination(s) of 2, 3, 4, 5, or more of the specific antigenic and/or immunogenic polypeptides disclosed herein. Further included in the present invention are antibodies which bind polypeptides encoded by polynucleotides which hybridize to a polynucleotide of the present invention under stringent hybridization conditions (as described herein). Antibodies of the present invention may also be described or specified in terms of their binding affinity to a polypeptide of the invention. Preferred binding affinities include those with a dissociation constant or Kd less than 5×10⁻² M, 10⁻² M, 5×10⁻³ M, 10⁻³ M, 5×10⁻⁴M, 10⁻⁴ M, 5×10⁻⁵ M, 10⁻⁵ M, 5×10⁻⁶ M, 10⁻⁶M, 5×10⁻⁷ M, 10⁷ M, 5×10⁻⁸ M, 10⁻⁸ M, 5×10⁻⁹ M, 10⁻⁹ M, 5×10⁻¹⁰ M, 10⁻¹⁰M, 5×10⁻¹¹ M, 10⁻¹¹ M, 5×10⁻¹² M, 10⁻¹² M, 5×10⁻¹³ M, 10⁻¹³ M, 5×10⁻¹⁴ M, 10⁻¹⁴ M, 5×10⁻¹⁵ M, or 10⁻¹⁵ M.

The invention also provides antibodies that competitively inhibit binding of an antibody to an epitope of the invention as determined by any method known in the art for determining competitive binding, for example, the immunoassays described herein. In preferred embodiments, the antibody competitively inhibits binding to the epitope by at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 60%, or at least 50%.

Antibodies of the present invention may act as agonists or antagonists of the polypeptides of the present invention. For example, the present invention includes antibodies which disrupt the receptor/ligand interactions with the polypeptides of the invention either partially or fully. Preferably, antibodies of the present invention bind an antigenic epitope disclosed herein, or a portion thereof. The invention features both receptor-specific antibodies and ligand-specific antibodies. The invention also features receptor-specific antibodies which do not prevent ligand binding but prevent receptor activation. Receptor activation (i.e., signaling) may be determined by techniques described herein or otherwise known in the art. For example, receptor activation can be determined by detecting the phosphorylation (e.g., tyrosine or serine/threonine) of the receptor or its substrate by immunoprecipitation followed by western blot analysis (for example, as described supra). In specific embodiments, antibodies are provided that inhibit ligand activity or receptor activity by at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 60%, or at least 50% of the activity in absence of the antibody.

The invention also features receptor-specific antibodies which both prevent ligand binding and receptor activation as well as antibodies that recognize the receptor-ligand complex, and, preferably, do not specifically recognize the unbound receptor or the unbound ligand. Likewise, included in the invention are neutralizing antibodies which bind the ligand and prevent binding of the ligand to the receptor, as well as antibodies which bind the ligand, thereby preventing receptor activation, but do not prevent the ligand from binding the receptor. Further included in the invention are antibodies which activate the receptor. These antibodies may act as receptor agonists, i.e., potentiate or activate either all or a subset of the biological activities of the ligand-mediated receptor activation, for example, by inducing dimerization of the receptor. The antibodies may be specified as agonists, antagonists or inverse agonists for biological activities comprising the specific biological activities of the peptides of the invention disclosed herein. The above antibody agonists can be made using methods known in the art. See, e.g., PCT publication WO 96/40281; U.S. Pat. No. 5,811,097; Deng et al., Blood 92(6):1981-1988 (1998); Chen et al., Cancer Res. 58(16):3668-3678 (1998); Harrop et al., J. Immunol. 161(4):1786-1794 (1998); Zhu et al., Cancer Res. 58(15):3209-3214 (1998); Yoon et al., J. Immunol. 160(7):3170-3179 (1998); Prat et al., J. Cell. Sci. 111(Pt2):237-247 (1998); Pitard et al., J. Immunol. Methods 205(2):177-190 (1997); Liautard et al., Cytokine 9(4):233-241 (1997); Carlson et al., J. Biol. Chem. 272(17):11295-11301 (1997); Taryman et al., Neuron 14(4):755-762 (1995); Muller et al., Structure 6(9):1153-1167 (1998); Bartunek et al., Cytokine 8(1):14-20 (1996) (which are all incorporated by reference herein in their entireties).

Antibodies of the present invention may be used, for example, to purify, detect, and target the polypeptides of the present invention, including both in vitro and in vivo diagnostic and therapeutic methods. For example, the antibodies have utility in immunoassays for qualitatively and quantitatively measuring levels of the polypeptides of the present invention in biological samples. See, e.g., Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); incorporated by reference herein in its entirety.

As discussed in more detail below, the antibodies of the present invention may be used either alone or in combination with other compositions. The antibodies may further be recombinantly fused to a heterologous polypeptide at the N- or C-terminus or chemically conjugated (including covalent and non-covalent conjugations) to polypeptides or other compositions. For example, antibodies of the present invention may be recombinantly fused or conjugated to molecules useful as labels in detection assays and effector molecules such as heterologous polypeptides, drugs, radionuclides, or toxins. See, e.g., PCT publications WO 92/08495; WO 91/14438; WO 89/12624; U.S. Pat. No. 5,314,995; and EP 396,387; the disclosures of which are incorporated herein by reference in their entireties.

The antibodies of the invention include derivatives that are modified, i.e, by the covalent attachment of any type of molecule to the antibody such that covalent attachment does not prevent the antibody from generating an anti-idiotypic response. For example, but not by way of limitation, the antibody derivatives include antibodies that have been modified, e.g., by glycosylation, acetylation, pegylation, phosphylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. Any of numerous chemical modifications may be carried out by known techniques, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Additionally, the derivative may contain one or more non-classical amino acids.

The antibodies of the present invention may be generated by any suitable method known in the art. Polyclonal antibodies to an antigen-of-interest can be produced by various procedures well known in the art. For example, a polypeptide of the invention can be administered to various host animals including, but not limited to, rabbits, mice, rats, etc. to induce the production of sera containing polyclonal antibodies specific for the antigen. Various adjuvants may be used to increase the immunological response, depending on the host species, and include but are not limited to, Freund's (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanins, dinitrophenol, and potentially useful human adjuvants such as BCG (bacille Calmette-Guerin) and corynebacterium parvum. Such adjuvants are also well known in the art.

Monoclonal antibodies can be prepared using a wide variety of techniques known in the art including the use of hybridoma, recombinant, and phage display technologies, or a combination thereof. For example, monoclonal antibodies can be produced using hybridoma techniques including those known in the art and taught, for example, in Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling, et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981) (said references incorporated by reference in their entireties). The term “monoclonal antibody” as used herein is not limited to antibodies produced through hybridoma technology. The term “monoclonal antibody” refers to an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced.

Methods for producing and screening for specific antibodies using hybridoma technology are routine and well known in the art and are discussed in detail in the Examples. In a non-limiting example, mice can be immunized with a polypeptide of the invention or a cell expressing such peptide. Once an immune response is detected, e.g., antibodies specific for the antigen are detected in the mouse serum, the mouse spleen is harvested and splenocytes isolated. The splenocytes are then fused by well known techniques to any suitable myeloma cells, for example cells from cell line SP20 available from the ATCC. Hybridomas are selected and cloned by limited dilution. The hybridoma clones are then assayed by methods known in the art for cells that secrete antibodies capable of binding a polypeptide of the invention. Ascites fluid, which generally contains high levels of antibodies, can be generated by immunizing mice with positive hybridoma clones.

Accordingly, the present invention provides methods of generating monoclonal antibodies as well as antibodies produced by the method comprising culturing a hybridoma cell secreting an antibody of the invention wherein, preferably, the hybridoma is generated by fusing splenocytes isolated from a mouse immunized with an antigen of the invention with myeloma cells and then screening the hybridomas resulting from the fusion for hybridoma clones that secrete an antibody able to bind a polypeptide of the invention.

Another well known method for producing both polyclonal and monoclonal human B cell lines is transformation using Epstein Barr Virus (EBV). Protocols for generating EBV-transformed B cell lines are commonly known in the art, such as, for example, the protocol outlined in Chapter 7.22 of Current Protocols in Immunology, Coligan et al., Eds., 1994, John Wiley & Sons, NY, which is hereby incorporated in its entirety by reference. The source of B cells for transformation is commonly human peripheral blood, but B cells for transformation may also be derived from other sources including, but not limited to, lymph nodes, tonsil, spleen, tumor tissue, and infected tissues. Tissues are generally made into single cell suspensions prior to EBV transformation. Additionally, steps may be taken to either physically remove or inactivate T cells (e.g., by treatment with cyclosporin A) in B cell-containing samples, because T cells from individuals seropositive for anti-EBV antibodies can suppress B cell immortalization by EBV.

In general, the sample containing human B cells is innoculated with EBV, and cultured for 3-4 weeks. A typical source of EBV is the culture supernatant of the B95-8 cell line (ATCC #VR-1492). Physical signs of EBV transformation can generally be seen towards the end of the 3-4 week culture period. By phase-contrast microscopy, transformed cells may appear large, clear, hairy and tend to aggregate in tight clusters of cells. Initially, EBV lines are generally polyclonal. However, over prolonged periods of cell cultures, EBV lines may become monoclonal or polyclonal as a result of the selective outgrowth of particular B cell clones. Alternatively, polyclonal EBV transformed lines may be subcloned (e.g., by limiting dilution culture) or fused with a suitable fusion partner and plated at limiting dilution to obtain monoclonal B cell lines. Suitable fusion partners for EBV transformed cell lines include mouse myeloma cell lines (e.g., SP2/0, X63-Ag8.653), heteromyeloma cell lines (human×mouse; e.g, SPAM-8, SBC-H20, and CB-F7), and human cell lines (e.g., GM 1500, SKO-007, RPMI 8226, and KR-4). Thus, the present invention also provides a method of generating polyclonal or monoclonal human antibodies against polypeptides of the invention or fragments thereof, comprising EBV-transformation of human B cells.

Antibody fragments which recognize specific epitopes may be generated by known techniques. For example, Fab and F(ab′)2 fragments of the invention may be produced by proteolytic cleavage of immunoglobulin molecules, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab′)2 fragments). F(ab′)2 fragments contain the variable region, the light chain constant region and the CH1 domain of the heavy chain.

For example, the antibodies of the present invention can also be generated using various phage display methods known in the art. In phage display methods, functional antibody domains are displayed on the surface of phage particles which carry the polynucleotide sequences encoding them. In a particular embodiment, such phage can be utilized to display antigen binding domains expressed from a repertoire or combinatorial antibody library (e.g., human or murine). Phage expressing an antigen binding domain that binds the antigen of interest can be selected or identified with antigen, e.g., using labeled antigen or antigen bound or captured to a solid surface or bead. Phage used in these methods are typically filamentous phage including fd and M13 binding domains expressed from phage with Fab, Fv or disulfide stabilized Fv antibody domains recombinantly fused to either the phage gene III or gene VIII protein. Examples of phage display methods that can be used to make the antibodies of the present invention include those disclosed in Brinkman et al., J. Immunol. Methods 182:41-50 (1995); Ames et al., J. Immunol. Methods 184:177-186 (1995); Kettleborough et al., Eur. J. Immunol. 24:952-958 (1994); Persic et al., Gene 187 9-18 (1997); Burton et al., Advances in Immunology 57:191-280 (1994); PCT application No. PCT/GB91/01134; PCT publications WO 90/02809; WO 91/10737; WO 92/01047; WO 92/18619; WO 93/11236; WO 95/15982; WO 95/20401; and U.S. Pat. Nos. 5,698,426; 5,223,409; 5,403,484; 5,580,717; 5,427,908; 5,750,753; 5,821,047; 5,571,698; 5,427,908; 5,516,637; 5,780,225; 5,658,727; 5,733,743 and 5,969,108; each of which is incorporated herein by reference in its entirety.

As described in the above references, after phage selection, the antibody coding regions from the phage can be isolated and used to generate whole antibodies, including human antibodies, or any other desired antigen binding fragment, and expressed in any desired host, including mammalian cells, insect cells, plant cells, yeast, and bacteria, e.g., as described in detail below. For example, techniques to recombinantly produce Fab, Fab′ and F(ab′)2 fragments can also be employed using methods known in the art such as those disclosed in PCT publication WO 92/22324; Mullinax et al., BioTechniques 12(6):864-869 (1992); and Sawai et al., AJRI 34:26-34 (1995); and Better et al., Science 240:1041-1043 (1988) (said references incorporated by reference in their entireties).

Examples of techniques which can be used to produce single-chain Fvs and antibodies include those described in U.S. Pat. Nos. 4,946,778 and 5,258,498; Huston et al., Methods in Enzymology 203:46-88 (1991); Shu et al., PNAS 90:7995-7999 (1993); and Skerra et al., Science 240:1038-1040 (1988). For some uses, including in vivo use of antibodies in humans and in vitro detection assays, it may be preferable to use chimeric, humanized, or human antibodies. A chimeric antibody is a molecule in which different portions of the antibody are derived from different animal species, such as antibodies having a variable region derived from a murine monoclonal antibody and a human immunoglobulin constant region. Methods for producing chimeric antibodies are known in the art. See e.g., Morrison, Science 229:1202 (1985); Oi et al., BioTechniques 4:214 (1986); Gillies et al., (1989) J. Immunol. Methods 125:191-202; U.S. Pat. Nos. 5,807,715; 4,816,567; and 4,816,397, which are incorporated herein by reference in their entirety. Humanized antibodies are antibody molecules from non-human species antibody that binds the desired antigen having one or more complementarity determining regions (CDRs) from the non-human species and a framework regions from a human immunoglobulin molecule. Often, framework residues in the human framework regions will be substituted with the corresponding residue from the CDR donor antibody to alter, preferably improve, antigen binding. These framework substitutions are identified by methods well known in the art, e.g., by modeling of the interactions of the CDR and framework residues to identify framework residues important for antigen binding and sequence comparison to identify unusual framework residues at particular positions. (See, e.g., Queen et al., U.S. Pat. No. 5,585,089; Riechmann et al., Nature 332:323 (1988), which are incorporated herein by reference in their entireties.) Antibodies can be humanized using a variety of techniques known in the art including, for example, CDR-grafting (EP 239,400; PCT publication WO 91/09967; U.S. Pat. Nos. 5,225,539; 5,530,101; and 5,585,089), veneering or resurfacing (EP 592,106; EP 519,596; Padlan, Molecular Immunology 28(4/5):489-498 (1991); Studnicka et al., Protein Engineering 7(6):805-814 (1994); Roguska. et al., PNAS 91:969-973 (1994)), and chain shuffling (U.S. Pat. No. 5,565,332).

Completely human antibodies are particularly desirable for therapeutic treatment of human patients. Human antibodies can be made by a variety of methods known in the art including phage display methods described above using antibody libraries derived from human immunoglobulin sequences. See also, U.S. Pat. Nos. 4,444,887 and 4,716,111; and PCT publications WO 98/46645, WO 98/50433, WO 98/24893, WO 98/16654, WO 96/34096, WO 96/33735, and WO 91/10741; each of which is incorporated herein by reference in its entirety.

Human antibodies can also be produced using transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which can express human immunoglobulin genes. For example, the human heavy and light chain immunoglobulin gene complexes may be introduced randomly or by homologous recombination into mouse embryonic stem cells. Alternatively, the human variable region, constant region, and diversity region may be introduced into mouse embryonic stem cells in addition to the human heavy and light chain genes. The mouse heavy and light chain immunoglobulin genes may be rendered non-functional separately or simultaneously with the introduction of human immunoglobulin loci by homologous recombination. In particular, homozygous deletion of the JH region prevents endogenous antibody production. The modified embryonic stem cells are expanded and microinjected into blastocysts to produce chimeric mice. The chimeric mice are then bred to produce homozygous offspring which express human antibodies. The transgenic mice are immunized in the normal fashion with a selected antigen, e.g., all or a portion of a polypeptide of the invention. Monoclonal antibodies directed against the antigen can be obtained from the immunized, transgenic mice using conventional hybridoma technology. The human immunoglobulin transgenes harbored by the transgenic mice rearrange during B cell differentiation, and subsequently undergo class switching and somatic mutation. Thus, using such a technique, it is possible to produce therapeutically useful IgG, IgA, IgM and IgE antibodies. For an overview of this technology for producing human antibodies, see Lonberg and Huszar, Int. Rev. Immunol. 13:65-93 (1995). For a detailed discussion of this technology for producing human antibodies and human monoclonal antibodies and protocols for producing such antibodies, see, e.g., PCT publications WO 98/24893; WO 92/01047; WO 96/34096; WO 96/33735; European Patent No. 0 598 877; U.S. Pat. Nos. 5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016; 5,545,806; 5,814,318; 5,885,793; 5,916,771; 5,939,598; 6,075,181; and 6,114,598, which are incorporated by reference herein in their entirety. In addition, companies such as Abgenix, Inc. (Freemont, Calif.) and Genpharm (San Jose, Calif.) can be engaged to provide human antibodies directed against a selected antigen using technology similar to that described above.

Completely human antibodies which recognize a selected epitope can be generated using a technique referred to as “guided selection.” In this approach a selected non-human monoclonal antibody, e.g., a mouse antibody, is used to guide the selection of a completely human antibody recognizing the same epitope. (Jespers et al., Bio/technology 12:899-903 (1988)).

Further, antibodies to the polypeptides of the invention can, in turn, be utilized to generate anti-idiotype antibodies that “mimic” polypeptides of the invention using techniques well known to those skilled in the art. (See, e.g., Greenspan & Bona, FASEB J. 7(5):437-444; (1989) and Nissinoff, J. Immunol. 147(8):2429-2438 (1991)). For example, antibodies which bind to and competitively inhibit polypeptide multimerization and/or binding of a polypeptide of the invention to a ligand can be used to generate anti-idiotypes that “mimic” the polypeptide multimerization and/or binding domain and, as a consequence, bind to and neutralize polypeptide and/or its ligand. Such neutralizing anti-idiotypes or Fab fragments of such anti-idiotypes can be used in therapeutic regimens to neutralize polypeptide ligand(s)/receptor(s). For example, such anti-idiotypic antibodies can be used to bind a polypeptide of the invention and/or to bind its ligand(s)/receptor(s), and thereby block its biological activity. Alternatively, antibodies which bind to and enhance polypeptide multimerization and/or binding, and/or receptor/ligand multimerization, binding and/or signaling can be used to generate anti-idiotypes that function as agonists of a polypeptide of the invention and/or its ligand/receptor. Such agonistic anti-idiotypes or Fab fragments of such anti-idiotypes can be used in therapeutic regimens as agonists of the polypeptides of the invention or its ligand(s)/receptor(s). For example, such anti-idiotypic antibodies can be used to bind a polypeptide of the invention and/or to bind its ligand(s)/receptor(s), and thereby promote or enhance its biological activity.

Intrabodies of the invention can be produced using methods known in the art, such as those disclosed and reviewed in Chen et al., Hum. Gene Ther. 5:595-601 (1994); Marasco, W. A., Gene Ther. 4:11-15 (1997); Rondon and Marasco, Annu. Rev. Microbiol. 51:257-283 (1997); Proba et al., J. Mol. Biol. 275:245-253 (1998); Cohen et al., Oncogene 17:2445-2456 (1998); Ohage and Steipe, J. Mol. Biol. 291:1119-1128 (1999); Ohage et al., J. Mol. Biol. 291:1129-1134 (1999); Wirtz and Steipe, Protein Sci. 8:2245-2250 (1999); Zhu et al., J. Immunol. Methods 231:207-222 (1999); and references cited therein.

Polynucleotides Encoding Antibodies

The invention further provides polynucleotides comprising a nucleotide sequence encoding an antibody of the invention and fragments thereof. The invention also encompasses polynucleotides that hybridize under stringent or alternatively, under lower stringency hybridization conditions, e.g., as defined supra, to polynucleotides that encode an antibody, preferably, that specifically binds to a polypeptide of the invention, preferably, an antibody that binds to a polypeptide having the amino acid sequence of SEQ ID NO:Y, to a polypeptide encoded by a portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, and/or to a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

The polynucleotides may be obtained, and the nucleotide sequence of the polynucleotides determined, by any method known in the art. For example, if the nucleotide sequence of the antibody is known, a polynucleotide encoding the antibody may be assembled from chemically synthesized oligonucleotides (e.g., as described in Kutmeier et al., BioTechniques 17:242 (1994)), which, briefly, involves the synthesis of overlapping oligonucleotides containing portions of the sequence encoding the antibody, annealing and ligating of those oligonucleotides, and then amplification of the ligated oligonucleotides by PCR.

Alternatively, a polynucleotide encoding an antibody may be generated from nucleic acid from a suitable source. If a clone containing a nucleic acid encoding a particular antibody is not available, but the sequence of the antibody molecule is known, a nucleic acid encoding the immunoglobulin may be chemically synthesized or obtained from a suitable source (e.g., an antibody cDNA library, or a cDNA library generated from, or nucleic acid, preferably poly A+ RNA, isolated from, any tissue or cells expressing the antibody, such as hybridoma cells selected to express an antibody of the invention) by PCR amplification using synthetic primers hybridizable to the 3′ and 5′ ends of the sequence or by cloning using an oligonucleotide probe specific for the particular gene sequence to identify, e.g., a cDNA clone from a cDNA library that encodes the antibody. Amplified nucleic acids generated by PCR may then be cloned into replicable cloning vectors using any method well known in the art.

Once the nucleotide sequence and corresponding amino acid sequence of the antibody is determined, the nucleotide sequence of the antibody may be manipulated using methods well known in the art for the manipulation of nucleotide sequences, e.g., recombinant DNA techniques, site directed mutagenesis, PCR, etc. (see, for example, the techniques described in Sambrook et al., 1990, Molecular Cloning, A Laboratory Manual, 2d Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. and Ausubel et al., eds., 1998, Current Protocols in Molecular Biology, John Wiley & Sons, NY, which are both incorporated by reference herein in their entireties), to generate antibodies having a different amino acid sequence, for example to create amino acid substitutions, deletions, and/or insertions.

In a specific embodiment, the amino acid sequence of the heavy and/or light chain variable domains may be inspected to identify the sequences of the complementarity determining regions (CDRs) by methods that are well know in the art, e.g., by comparison to known amino acid sequences of other heavy and light chain variable regions to determine the regions of sequence hypervariability. Using routine recombinant DNA techniques, one or more of the CDRs may be inserted within framework regions, e.g., into human framework regions to humanize a non-human antibody, as described supra. The framework regions may be naturally occurring or consensus framework regions, and preferably human framework regions (see, e.g., Chothia et al., J. Mol. Biol. 278: 457-479 (1998) for a listing of human framework regions). Preferably, the polynucleotide generated by the combination of the framework regions and CDRs encodes an antibody that specifically binds a polypeptide of the invention. Preferably, as discussed supra, one or more amino acid substitutions may be made within the framework regions, and, preferably, the amino acid substitutions improve binding of the antibody to its antigen. Additionally, such methods may be used to make amino acid substitutions or deletions of one or more variable region cysteine residues participating in an intrachain disulfide bond to generate antibody molecules lacking one or more intrachain disulfide bonds. Other alterations to the polynucleotide are encompassed by the present invention and within the skill of the art.

In addition, techniques developed for the production of “chimeric antibodies” (Morrison et al., Proc. Natl. Acad. Sci. 81:851-855 (1984); Neuberger et al., Nature 312:604-608 (1984); Takeda et al., Nature 314:452-454 (1985)) by splicing genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity can be used. As described supra, a chimeric antibody is a molecule in which different portions are derived from different animal species, such as those having a variable region derived from a murine mAb and a human immunoglobulin constant region, e.g., humanized antibodies.

Alternatively, techniques described for the production of single chain antibodies (U.S. Pat. No. 4,946,778; Bird, Science 242:423-42 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Ward et al., Nature 334:544-54 (1989)) can be adapted to produce single chain antibodies. Single chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single chain polypeptide. Techniques for the assembly of functional Fv fragments in E. coli may also be used (Skerra et al., Science 242:1038-1041 (1988)).

Methods of Producing Antibodies

The antibodies of the invention can be produced by any method known in the art for the synthesis of antibodies, in particular, by chemical synthesis or preferably, by recombinant expression techniques. Methods of producing antibodies include, but are not limited to, hybridoma technology, EBV transformation, and other methods discussed herein as well as through the use recombinant DNA technology, as discussed below.

Recombinant expression of an antibody of the invention, or fragment, derivative or analog thereof, (e.g., a heavy or light chain of an antibody of the invention or a single chain antibody of the invention), requires construction of an expression vector containing a polynucleotide that encodes the antibody. Once a polynucleotide encoding an antibody molecule or a heavy or light chain of an antibody, or portion thereof (preferably containing the heavy or light chain variable domain), of the invention has been obtained, the vector for the production of the antibody molecule may be produced by recombinant DNA technology using techniques well known in the art. Thus, methods for preparing a protein by expressing a polynucleotide containing an antibody encoding nucleotide sequence are described herein. Methods which are well known to those skilled in the art can be used to construct expression vectors containing antibody coding sequences and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. The invention, thus, provides replicable vectors comprising a nucleotide sequence encoding an antibody molecule of the invention, or a heavy or light chain thereof, or a heavy or light chain variable domain, operably linked to a promoter. Such vectors may include the nucleotide sequence encoding the constant region of the antibody molecule (see, e.g., PCT Publication WO 86/05807; PCT Publication WO 89/01036; and U.S. Pat. No. 5,122,464) and the variable domain of the antibody may be cloned into such a vector for expression of the entire heavy or light chain.

The expression vector is transferred to a host cell by conventional techniques and the transfected cells are then cultured by conventional techniques to produce an antibody of the invention. Thus, the invention includes host cells containing a polynucleotide encoding an antibody of the invention, or a heavy or light chain thereof, or a single chain antibody of the invention, operably linked to a heterologous promoter. In preferred embodiments for the expression of double-chained antibodies, vectors encoding both the heavy and light chains may be co-expressed in the host cell for expression of the entire immunoglobulin molecule, as detailed below.

A variety of host-expression vector systems may be utilized to express the antibody molecules of the invention. Such host-expression systems represent vehicles by which the coding sequences of interest may be produced and subsequently purified, but also represent cells which may, when transformed or transfected with the appropriate nucleotide coding sequences, express an antibody molecule of the invention in situ. These include but are not limited to microorganisms such as bacteria (e.g., E. coli, B. subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing antibody coding sequences; yeast (e.g., Saccharomyces, Pichia) transformed with recombinant yeast expression vectors containing antibody coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing antibody coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing antibody coding sequences; or mammalian cell systems (e.g., COS, CHO, BHK, 293, 3T3 cells) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter). Preferably, bacterial cells such as Escherichia coli, and more preferably, eukaryotic cells, especially for the expression of whole recombinant antibody molecule, are used for the expression of a recombinant antibody molecule. For example, mammalian cells such as Chinese hamster ovary cells (CHO), in conjunction with a vector such as the major intermediate early gene promoter element from human cytomegalovirus is an effective expression system for antibodies (Foecking et al., Gene 45:101 (1986); Cockett et al., Bio/Technology 8:2 (1990)).

In bacterial systems, a number of expression vectors may be advantageously selected depending upon the use intended for the antibody molecule being expressed. For example, when a large quantity of such a protein is to be produced, for the generation of pharmaceutical compositions of an antibody molecule, vectors which direct the expression of high levels of fusion protein products that are readily purified may be desirable. Such vectors include, but are not limited, to the E. coli expression vector pUR278 (Ruther et al., EMBO J. 2:1791 (1983)), in which the antibody coding sequence may be ligated individually into the vector in frame with the lac Z coding region so that a fusion protein is produced; pIN vectors (Inouye & Inouye, Nucleic Acids Res. 13:3101-3109 (1985); Van Heeke & Schuster, J. Biol. Chem. 24:5503-5509 (1989)); and the like. pGEX vectors may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption and binding to matrix glutathione-agarose beads followed by elution in the presence of free glutathione. The pGEX vectors are designed to include thrombin or factor Xa protease cleavage sites so that the cloned target gene product can be released from the GST moiety.

In an insect system, Autographa californica nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes. The virus grows in Spodoptera frugiperda cells. The antibody coding sequence may be cloned individually into non-essential regions (for example the polyhedrin gene) of the virus and placed under control of an AcNPV promoter (for example the polyhedrin promoter).

In mammalian host cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, the antibody coding sequence of interest may be ligated to an adenovirus transcription/translation control complex, e.g., the late promoter and tripartite leader sequence. This chimeric gene may then be inserted in the adenovirus genome by in vitro or in vivo recombination. Insertion in a non-essential region of the viral genome (e.g., region E1 or E3) will result in a recombinant virus that is viable and capable of expressing the antibody molecule in infected hosts. (e.g., see Logan & Shenk, Proc. Natl. Acad. Sci. USA 81:355-359 (1984)). Specific initiation signals may also be required for efficient translation of inserted antibody coding sequences. These signals include the ATG initiation codon and adjacent sequences. Furthermore, the initiation codon must be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert. These exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of appropriate transcription enhancer elements, transcription terminators, etc. (see Bittner et al., Methods in Enzymol. 153:51-544 (1987)).

In addition, a host cell strain may be chosen which modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific fashion desired. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may be important for the function of the protein. Different host cells have characteristic and specific mechanisms for the post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the foreign protein expressed. To this end, eukaryotic host cells which possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product may be used. Such mammalian host cells include but are not limited to CHO, VERY, BHK, Hela, COS, MDCK, 293, 3T3, W138, and in particular, breast cancer cell lines such as, for example, BT483, Hs578T, HTB2, BT20 and T47D, and normal mammary gland cell line such as, for example, CRL7030 and Hs578Bst.

For long-term, high-yield production of recombinant proteins, stable expression is preferred. For example, cell lines which stably express the antibody molecule may be engineered. Rather than using expression vectors which contain viral origins of replication, host cells can be transformed with DNA controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of the foreign DNA, engineered cells may be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci which in turn can be cloned and expanded into cell lines. This method may advantageously be used to engineer cell lines which express the antibody molecule. Such engineered cell lines may be particularly useful in screening and evaluation of compounds that interact directly or indirectly with the antibody molecule.

A number of selection systems may be used, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al., Cell 11:223 (1977)), hypoxanthine-guanine phosphoribosyltransferase (Szybalska & Szybalski, Proc. Natl. Acad. Sci. USA 48:202 (1992)), and adenine phosphoribosyltransferase (Lowy et al., Cell 22:817 (1980)) genes can be employed in tk−, hgprt− or aprt− cells, respectively. Also, antimetabolite resistance can be used as the basis of selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler et al., Natl. Acad. Sci. USA 77:357 (1980); O'Hare et al., Proc. Natl. Acad. Sci. USA 78:1527 (1981)); gpt, which confers resistance to mycophenolic acid (Mulligan & Berg, Proc. Natl. Acad. Sci. USA 78:2072 (1981)); neo, which confers resistance to the aminoglycoside G-418 Clinical Pharmacy 12:488-505; Wu and Wu, Biotherapy 3:87-95 (1991); Tolstoshev, Ann. Rev. Pharmacol. Toxicol. 32:573-596 (1993); Mulligan, Science 260:926-932 (1993); and Morgan and Anderson, Ann. Rev. Biochem. 62:191-217 (1993); May, 1993, TIB TECH 11(5):155-215 (1993)); and hygro, which confers resistance to hygromycin (Santerre et al., Gene 30:147 (1984)). Methods commonly known in the art of recombinant DNA technology may be routinely applied to select the desired recombinant clone, and such methods are described, for example, in Ausubel et al. (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993); Kriegler, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990); and in Chapters 12 and 13, Dracopoli et al. (eds), Current Protocols in Human Genetics, John Wiley & Sons, NY (1994); Colberre-Garapin et al., J. Mol. Biol. 150:1 (1981), which are incorporated by reference herein in their entireties.

The expression levels of an antibody molecule can be increased by vector amplification (for a review, see Bebbington and Hentschel, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol. 3. (Academic Press, New York, 1987)). When a marker in the vector system expressing antibody is amplifiable, increase in the level of inhibitor present in culture of host cell will increase the number of copies of the marker gene. Since the amplified region is associated with the antibody gene, production of the antibody will also increase (Crouse et al., Mol. Cell. Biol. 3:257 (1983)).

Vectors which use glutamine synthase (GS) or DHFR as the selectable markers can be amplified in the presence of the drugs methionine sulphoximine or methotrexate, respectively. An advantage of glutamine synthase based vectors are the availability of cell lines (e.g., the murine myeloma cell line, NS0) which are glutamine synthase negative. Glutamine synthase expression systems can also function in glutamine synthase expressing cells (e.g. Chinese Hamster Ovary (CHO) cells) by providing additional inhibitor to prevent the functioning of the endogenous gene. A glutamine synthase expression system and components thereof are detailed in PCT publications: WO87/04462; WO86/05807; WO89/01036; WO89/10404; and WO91/06657 which are incorporated in their entireties by reference herein. Additionally, glutamine synthase expression vectors that may be used according to the present invention are commercially available from suppliers, including, for example Lonza Biologics, Inc. (Portsmouth, N.H.). Expression and production of monoclonal antibodies using a GS expression system in murine myeloma cells is described in Bebbington et al., Bio/technology 10:169(1992) and in Biblia and Robinson Biotechnol. Prog. 11:1 (1995) which are incorporated in their entireties by reference herein.

The host cell may be co-transfected with two expression vectors of the invention, the first vector encoding a heavy chain derived polypeptide and the second vector encoding a light chain derived polypeptide. The two vectors may contain identical selectable markers which enable equal expression of heavy and light chain polypeptides. Alternatively, a single vector may be used which encodes, and is capable of expressing, both heavy and light chain polypeptides. In such situations, the light chain should be placed before the heavy chain to avoid an excess of toxic free heavy chain (Proudfoot, Nature 322:52 (1986); Kohler, Proc. Natl. Acad. Sci. USA 77:2197 (1980)). The coding sequences for the heavy and light chains may comprise cDNA or genomic DNA.

Once an antibody molecule of the invention has been produced by an animal, chemically synthesized, or recombinantly expressed, it may be purified by any method known in the art for purification of an immunoglobulin molecule, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for the purification of proteins. In addition, the antibodies of the present invention or fragments thereof can be fused to heterologous polypeptide sequences described herein or otherwise known in the art, to facilitate purification.

The present invention encompasses antibodies recombinantly fused or chemically conjugated (including both covalently and non-covalently conjugations) to a polypeptide (or portion thereof, preferably at least 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 amino acids of the polypeptide) of the present invention to generate fusion proteins. The fusion does not necessarily need to be direct, but may occur through linker sequences. The antibodies may be specific for antigens other than polypeptides (or portion thereof, preferably at least 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 amino acids of the polypeptide) of the present invention. For example, antibodies may be used to target the polypeptides of the present invention to particular cell types, either in vitro or in vivo, by fusing or conjugating the polypeptides of the present invention to antibodies specific for particular cell surface receptors. Antibodies fused or conjugated to the polypeptides of the present invention may also be used in in vitro immunoassays and purification methods using methods known in the art. See e.g., Harbor et al., supra, and PCT publication WO 93/21232; EP 439,095; Naramura et al., Immunol. Lett. 39:91-99 (1994); U.S. Pat. No. 5,474,981; Gillies et al., PNAS 89:1428-1432 (1992); Fell et al., J. Immunol. 146:2446-2452 (1991), which are incorporated by reference in their entireties.

The present invention further includes compositions comprising the polypeptides of the present invention fused or conjugated to antibody domains other than the variable regions. For example, the polypeptides of the present invention may be fused or conjugated to an antibody Fc region, or portion thereof. The antibody portion fused to a polypeptide of the present invention may comprise the constant region, hinge region, CH1 domain, CH2 domain, and CH3 domain or any combination of whole domains or portions thereof. The polypeptides may also be fused or conjugated to the above antibody portions to form multimers. For example, Fc portions fused to the polypeptides of the present invention can form dimers through disulfide bonding between the Fc portions. Higher multimeric forms can be made by fusing the polypeptides to portions of IgA and IgM. Methods for fusing or conjugating the polypeptides of the present invention to antibody portions are known in the art. See, e.g., U.S. Pat. Nos. 5,336,603; 5,622,929; 5,359,046; 5,349,053; 5,447,851; 5,112,946; EP 307,434; EP 367,166; PCT publications WO 96/04388; WO 91/06570; Ashkenazi et al., Proc. Natl. Acad. Sci. USA 88:10535-10539 (1991); Zheng et al., J. Immunol. 154:5590-5600 (1995); and Vil et al., Proc. Natl. Acad. Sci. USA 89:11337-11341 (1992) (said references incorporated by reference in their entireties).

As discussed, supra, the polypeptides corresponding to a polypeptide, polypeptide fragment, or a variant of SEQ ID NO:Y may be fused or conjugated to the above antibody portions to increase the in vivo half life of the polypeptides or for use in immunoassays using methods known in the art. Further, the polypeptides corresponding to SEQ ID NO:Y may be fused or conjugated to the above antibody portions to facilitate purification. One reported example describes chimeric proteins consisting of the first two domains of the human CD4-polypeptide and various domains of the constant regions of the heavy or light chains of mammalian immunoglobulins. See EP 394,827; and Traunecker et al., Nature 331:84-86 (1988). The polypeptides of the present invention fused or conjugated to an antibody having disulfide-linked dimeric structures (due to the IgG) may also be more efficient in binding and neutralizing other molecules, than the monomeric secreted protein or protein fragment alone. See, for example, Fountoulakis et al., J. Biochem. 270:3958-3964 (1995). In many cases, the Fc part in a fusion protein is beneficial in therapy and diagnosis, and thus can result in, for example, improved pharmacokinetic properties. See, for example, EP A 232,262. Alternatively, deleting the Fc part after the fusion protein has been expressed, detected, and purified, would be desired. For example, the Fc portion may hinder therapy and diagnosis if the fusion protein is used as an antigen for immunizations. In drug discovery, for example, human proteins, such as hIL-5, have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. (See, Bennett et al., J. Molecular Recognition 8:52-58 (1995); Johanson et al., J. Biol. Chem. 270:9459-9471 (1995)).

Moreover, the antibodies or fragments thereof of the present invention can be fused to marker sequences, such as a peptide to facilitate purification. In preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, Calif., 91311), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. Other peptide tags useful for purification include, but are not limited to, the “HA” tag, which corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., Cell 37:767 (1984)) and the “flag” tag.

The present invention further encompasses antibodies or fragments thereof conjugated to a diagnostic or therapeutic agent. The antibodies can be used diagnostically to, for example, monitor the development or progression of a tumor as part of a clinical testing procedure to, e.g., determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron emitting metals using various positron emission tomographies, and nonradioactive paramagnetic metal ions. The detectable substance may be coupled or conjugated either directly to the antibody (or fragment thereof) or indirectly, through an intermediate (such as, for example, a linker known in the art) using techniques known in the art. See, for example, U.S. Pat. No. 4,741,900 for metal ions which can be conjugated to antibodies for use as diagnostics according to the present invention. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin; and examples of suitable radioactive material include 125I, 131I, 111In or 99Tc.

Further, an antibody or fragment thereof may be conjugated to a therapeutic moiety such as a cytotoxin, e.g., a cytostatic or cytocidal agent, a therapeutic agent or a radioactive metal ion, e.g., alpha-emitters such as, for example, 213Bi. A cytotoxin or cytotoxic agent includes any agent that is detrimental to cells. Examples include paclitaxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof. Therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents (e.g., vincristine and vinblastine).

The conjugates of the invention can be used for modifying a given biological response, the therapeutic agent or drug moiety is not to be construed as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor, α-interferon, β-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator, an apoptotic agent, e.g., TNF-alpha, TNF-beta, AIM I (See, International Publication No. WO 97/33899), AIM II (See, International Publication No. WO 97/34911), Fas Ligand (Takahashi et al., Int. Immunol., 6:1567-1574 (1994)), VEGI (See, International Publication No. WO 99/23105), a thrombotic agent or an anti-angiogenic agent, e.g., angiostatin or endostatin; or, biological response modifiers such as, for example, lymphokines, interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”), or other growth factors.

Antibodies may also be attached to solid supports, which are particularly useful for immunoassays or purification of the target antigen. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene.

Techniques for conjugating such therapeutic moiety to antibodies are well known. See, for example, Amon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery”, in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); “Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., “The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates”, Immunol. Rev. 62:119-58 (1982).

Alternatively, an antibody can be conjugated to a second antibody to form an antibody heteroconjugate as described by Segal in U.S. Pat. No. 4,676,980, which is incorporated herein by reference in its entirety.

An antibody, with or without a therapeutic moiety conjugated to it, administered alone or in combination with cytotoxic factor(s) and/or cytokine(s) can be used as a therapeutic.

Immunophenotyping

The antibodies of the invention may be utilized for immunophenotyping of cell lines and biological samples. Translation products of the gene of the present invention may be useful as cell-specific markers, or more specifically as cellular markers that are differentially expressed at various stages of differentiation and/or maturation of particular cell types. Monoclonal antibodies directed against a specific epitope, or combination of epitopes, will allow for the screening of cellular populations expressing the marker. Various techniques can be utilized using monoclonal antibodies to screen for cellular populations expressing the marker(s), and include magnetic separation using antibody-coated magnetic beads, “panning” with antibody attached to a solid matrix (i.e., plate), and flow cytometry (See, e.g., U.S. Pat. No. 5,985,660; and Morrison et al., Cell, 96:737-49 (1999)).

These techniques allow for the screening of particular populations of cells, such as might be found with hematological malignancies (i.e. minimal residual disease (MRD) in acute leukemic patients) and “non-self” cells in transplantations to prevent Graft-versus-Host Disease (GVHD). Alternatively, these techniques allow for the screening of hematopoietic stem and progenitor cells capable of undergoing proliferation and/or differentiation, as might be found in human umbilical cord blood.

Assays for Antibody Binding

The antibodies of the invention may be assayed for immunospecific binding by any method known in the art. The immunoassays which can be used include but are not limited to competitive and non-competitive assay systems using techniques such as western blots, radioimmunoassays, ELISA (enzyme linked immunosorbent assay), “sandwich” immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, and protein A immunoassays, to name but a few. Such assays are routine and well known in the art (see, e.g., Ausubel et al, eds, 1994, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, which is incorporated by reference herein in its entirety). Exemplary immunoassays are described briefly below (but are not intended by way of limitation).

Immunoprecipitation protocols generally comprise lysing a population of cells in a lysis buffer such as RIPA buffer (1% NP-40 or Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 0.15 M NaCl, 0.01 M sodium phosphate at pH 7.2, 1% Trasylol) supplemented with protein phosphatase and/or protease inhibitors (e.g., EDTA, PMSF, aprotinin, sodium vanadate), adding the antibody of interest to the cell lysate, incubating for a period of time (e.g., 1-4 hours) at 4° C., adding protein A and/or protein G sepharose beads to the cell lysate, incubating for about an hour or more at 4° C., washing the beads in lysis buffer and resuspending the beads in SDS/sample buffer. The ability of the antibody of interest to immunoprecipitate a particular antigen can be assessed by, e.g., western blot analysis. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the binding of the antibody to an antigen and decrease the background (e.g., pre-clearing the cell lysate with sepharose beads). For further discussion regarding immunoprecipitation protocols see, e.g., Ausubel et al., eds., (1994), Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, section 10.16.1.

Western blot analysis generally comprises preparing protein samples, electrophoresis of the protein samples in a polyacrylamide gel (e.g., 8%-20% SDS-PAGE depending on the molecular weight of the antigen), transferring the protein sample from the polyacrylamide gel to a membrane such as nitrocellulose, PVDF or nylon, blocking the membrane in blocking solution (e.g., PBS with 3% BSA or non-fat milk), washing the membrane in washing buffer (e.g., PBS-Tween 20), blocking the membrane with primary antibody (the antibody of interest) diluted in blocking buffer, washing the membrane in washing buffer, blocking the membrane with a secondary antibody (which recognizes the primary antibody, e.g., an anti-human antibody) conjugated to an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) or radioactive molecule (e.g., 32P or 125I) diluted in blocking buffer, washing the membrane in wash buffer, and detecting the presence of the antigen. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected and to reduce the background noise. For further discussion regarding western blot protocols see, e.g., Ausubel et al, eds, (1994), Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, section 10.8.1.

ELISAs comprise preparing antigen, coating the well of a 96 well microtiter plate with the antigen, adding the antibody of interest conjugated to a detectable compound such as an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) to the well and incubating for a period of time, and detecting the presence of the antigen. In ELISAs the antibody of interest does not have to be conjugated to a detectable compound; instead, a second antibody (which recognizes the antibody of interest) conjugated to a detectable compound may be added to the well. Further, instead of coating the well with the antigen, the antibody may be coated to the well. In this case, a second antibody conjugated to a detectable compound may be added following the addition of the antigen of interest to the coated well. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected as well as other variations of ELISAs known in the art. For further discussion regarding ELISAs see, e.g., Ausubel et al, eds, (1994), Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, section 11.2.1.

The binding affinity of an antibody to an antigen and the off-rate of an antibody-antigen interaction can be determined by competitive binding assays. One example of a competitive binding assay is a radioimmunoassay comprising the incubation of labeled antigen (e.g., 3H or 125I) with the antibody of interest in the presence of increasing amounts of unlabeled antigen, and the detection of the antibody bound to the labeled antigen. The affinity of the antibody of interest for a particular antigen and the binding off-rates can be determined from the data by scatchard plot analysis. Competition with a second antibody can also be determined using radioimmunoassays. In this case, the antigen is incubated with antibody of interest conjugated to a labeled compound (e.g., 3H or 125I) in the presence of increasing amounts of an unlabeled second antibody.

Antibodies of the invention may be characterized using immunocytochemisty methods on cells (e.g., mammalian cells, such as CHO cells) transfected with a vector enabling the expression of an antigen or with vector alone using techniques commonly known in the art. Antibodies that bind antigen transfected cells, but not vector-only transfected cells, are antigen specific.

Therapeutic Uses

Table 1D: In preferred embodiments, the present invention encompasses a method of treating a disease or disorder listed in the “Preferred Indications” column of Table 1D; comprising administering to a patient in which such treatment, prevention, or amelioration is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) represented by Table 1A and Table 1D (in the same row as the disease or disorder to be treated is listed in the “Preferred Indications” column of Table 1D) in an amount effective to treat, prevent, or ameliorate the disease or disorder.

As indicated in Table 1D, the polynucleotides, polypeptides, agonists, or antagonists of the present invention (including antibodies) can be used in assays to test for one or more biological activities. If these polynucleotides and polypeptides do exhibit activity in a particular assay, it is likely that these molecules may be involved in the diseases associated with the biological activity. Thus, the polynucleotides or polypeptides, or agonists or antagonists thereof (including antibodies) could be used to treat the associated disease.

The present invention encompasses methods of preventing, treating, diagnosing, or ameliorating a disease or disorder. In preferred embodiments, the present invention encompasses a method of treating a disease or disorder listed in the “Preferred Indications” column of Table 1D; comprising administering to a patient in which such treatment, prevention, or amelioration is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to treat, prevent, diagnose, or ameliorate the disease or disorder. The first and second columns of Table 1D show the “Gene No.” and “cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may be used in preventing, treating, diagnosing, or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in Column 3 of Table 1D.

In another embodiment, the present invention also encompasses methods of preventing, treating, diagnosing, or ameliorating a disease or disorder listed in the “Preferred Indications” column of Table 1D; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in Column 3 of Table 1D.

The “Preferred Indication” column describes diseases, disorders, and/or conditions that may be treated, prevented, diagnosed, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).

The recitation of “Cancer” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., leukemias, cancers, and/or as described below under “Hyperproliferative Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D may be used for example, to diagnose, treat, prevent, and/or ameliorate a neoplasm located in a tissue selected from the group consisting of: colon, abdomen, bone, breast, digestive system, liver, pancreas, prostate, peritoneum, lung, blood (e.g., leukemia), endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid), uterus, eye, head and neck, nervous (central and peripheral), lymphatic system, pelvic, skin, soft tissue, spleen, thoracic, and urogenital.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a pre-neoplastic condition, selected from the group consisting of: hyperplasia (e.g., endometrial hyperplasia and/or as described in the section entitled “Hyperproliferative Disorders”), metaplasia (e.g., connective tissue metaplasia, atypical metaplasia, and/or as described in the section entitled “Hyperproliferative Disorders”), and/or dysplasia (e.g., cervical dysplasia, and bronchopulmonary dysplasia).

In another specific embodiment, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cancer” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a benign dysproliferative disorder selected from the group consisting of: benign tumors, fibrocystic conditions, tissue hypertrophy, and/or as described in the section entitled “Hyperproliferative Disorders”.

The recitation of “Immune/Hematopoietic” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), blood disorders (e.g., as described below under “Immune Activity” “Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under “Infectious Disease”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having the “Immune/Hematopoietic” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: anemia, pancytopenia, leukopenia, thrombocytopenia, leukemias, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphocytic anemia (ALL), plasmacytomas, multiple myeloma, Burkitt's lymphoma, arthritis, asthma, AIDS, autoimmune disease, rheumatoid arthritis, granulomatous disease, immune deficiency, inflammatory bowel disease, sepsis, neutropenia, neutrophilia, psoriasis, immune reactions to transplanted organs and tissues, systemic lupus erythematosis, hemophilia, hypercoagulation, diabetes mellitus, endocarditis, meningitis, Lyme Disease, and allergies.

The recitation of “Reproductive” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the reproductive system (e.g., as described below under “Reproductive System Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Reproductive” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cryptorchism, prostatitis, inguinal hernia, varicocele, leydig cell tumors, verrucous carcinoma, prostatitis, malacoplakia, Peyronie's disease, penile carcinoma, squamous cell hyperplasia, dysmenorrhea, ovarian adenocarcinoma, Turner's syndrome, mucopurulent cervicitis, Sertoli-leydig tumors, ovarian cancer, uterine cancer, pelvic inflammatory disease, testicular cancer, prostate cancer, Klinefelter's syndrome, Young's syndrome, premature ejaculation, diabetes mellitus, cystic fibrosis, Kartagener's syndrome, testicular atrophy, testicular feminization, anorchia, ectopic testis, epididymitis, orchitis, gonorrhea, syphilis, testicular torsion, vasitis nodosa, germ cell tumors, stromal tumors, dysmenorrhea, retroverted uterus, endometriosis, fibroids, adenomyosis, anovulatory bleeding, amenorrhea, Cushing's syndrome, hydatidiform moles, Asherman's syndrome, premature menopause, precocious puberty, uterine polyps, dysfunctional uterine bleeding, cervicitis, chronic cervicitis, mucopurulent cervicitis, cervical dysplasia, cervical polyps, Nabothian cysts, cervical erosion, cervical incompetence, cervical neoplasms, pseudohermaphroditism, and premenstrual syndrome.

The recitation of “Musculoskeletal” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the immune system (e.g., as described below under “Immune Activity”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Musculoskeletal” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: bone cancers (e.g., osteochondromas, benign chondromas, chondroblastoma, chondromyxoid fibromas, osteoid osteomas, giant cell tumors, multiple myeloma, osteosarcomas), Paget's Disease, rheumatoid arthritis, systemic lupus erythematosus, osteomyelitis, Lyme Disease, gout, bursitis, tendonitis, osteoporosis, osteoarthritis, muscular dystrophy, mitochondrial myopathy, cachexia, and multiple sclerosis.

The recitation of “Cardiovascular” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), and disorders of the cardiovascular system (e.g., as described below under “Cardiovascular Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Cardiovascular” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: myxomas, fibromas, rhabdomyomas, cardiovascular abnormalities (e.g., congenital heart defects, cerebral arteriovenous malformations, septal defects), heart disease (e.g., heart failure, congestive heart disease, arrhythmia, tachycardia, fibrillation, pericardial Disease, endocarditis), cardiac arrest, heart valve disease (e.g., stenosis, regurgitation, prolapse), vascular disease (e.g., hypertension, coronary artery disease, angina, aneurysm, arteriosclerosis, peripheral vascular disease), hyponatremia, hypernatremia, hypokalemia, and hyperkalemia.

The recitation of “Mixed Fetal” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferati ve Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Mixed Fetal” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: spina bifida, hydranencephaly, neurofibromatosis, fetal alcohol syndrome, diabetes mellitus, PKU, Down's syndrome, Patau syndrome, Edwards syndrome, Turner syndrome, Apert syndrome, Carpenter syndrome, Conradi syndrome, Crouzon syndrome, cutis laxa, Cornelia de Lange syndrome, Ellis-van Creveld syndrome, Holt-Oram syndrome, Kartagener syndrome, Meckel-Gruber syndrome, Noonan syndrome, Pallister-Hall syndrome, Rubinstein-Taybi syndrome, Scimitar syndrome, Smith-Lemli-Opitz syndrome, thromocytopenia-absent radius (TAR) syndrome, Treacher Collins syndrome, Williams syndrome, Hirschsprung's disease, Meckel's diverticulum, polycystic kidney disease, Turner's syndrome, and gonadal dysgenesis, Klippel-Feil syndrome, Ostogenesis imperfecta, muscular dystrophy, Tay-Sachs disease, Wilm's tumor, neuroblastoma, and retinoblastoma.

The recitation of “Excretory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and renal disorders (e.g., as described below under “Renal Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Excretory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: bladder cancer, prostate cancer, benign prostatic hyperplasia, bladder disorders (e.g., urinary incontinence, urinary retention, urinary obstruction, urinary tract Infections, interstitial cystitis, prostatitis, neurogenic bladder, hematuria), renal disorders (e.g., hydronephrosis, proteinuria, renal failure, pyelonephritis, urolithiasis, reflux nephropathy, and unilateral obstructive uropathy).

The recitation of “Neural/Sensory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the nervous system (e.g., as described below under “Neural Activity and Neurological Diseases”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Neural/Sensory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: brain cancer (e.g., brain stem glioma, brain tumors, central nervous system (Primary) lymphoma, central nervous system lymphoma, cerebellar astrocytoma, and cerebral astrocytoma, neurodegenerative disorders (e.g., Alzheimer's Disease, Creutzfeldt-Jakob Disease, Parkinson's Disease, and Idiopathic Presenile Dementia), encephalomyelitis, cerebral malaria, meningitis, metabolic brain diseases (e.g., phenylketonuria and pyruvate carboxylase deficiency), cerebellar ataxia, ataxia telangiectasia, and AIDS Dementia Complex, schizophrenia, attention deficit disorder, hyperactive attention deficit disorder, autism, and obsessive compulsive disorders.

The recitation of “Respiratory” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the respiratory system (e.g., as described below under “Respiratory Disorders”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Respiratory” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cancers of the respiratory system such as larynx cancer, pharynx cancer, trachea cancer, epiglottis cancer, lung cancer, squamous cell carcinomas, small cell (oat cell) carcinomas, large cell carcinomas, and adenocarcinomas. Allergic reactions, cystic fibrosis, sarcoidosis, histiocytosis X, infiltrative lung diseases (e.g., pulmonary fibrosis and lymphoid interstitial pneumonia), obstructive airway diseases (e.g., asthma, emphysema, chronic or acute bronchitis), occupational lung diseases (e.g., silicosis and asbestosis), pneumonia, and pleurisy.

The recitation of “Endocrine” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the respiratory system (e.g., as described below under “Respiratory Disorders”), renal disorders (e.g., as described below under “Renal Disorders”), and disorders of the endocrine system (e.g., as described below under “Endocrine Disorders”.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having an “Endocrine” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: cancers of endocrine tissues and organs (e.g., cancers of the hypothalamus, pituitary gland, thyroid gland, parathyroid glands, pancreas, adrenal glands, ovaries, and testes), diabetes (e.g., diabetes insipidus, type I and type II diabetes mellitus), obesity, disorders related to pituitary glands (e.g., hyperpituitarism, hypopituitarism, and pituitary dwarfism), hypothyroidism, hyperthyroidism, goiter, reproductive disorders (e.g. male and female infertility), disorders related to adrenal glands (e.g., Addison's Disease, corticosteroid deficiency, and Cushing's Syndrome), kidney cancer (e.g., hypernephroma, transitional cell cancer, and Wilm's tumor), diabetic nephropathy, interstitial nephritis, polycystic kidney disease, glomerulonephritis (e.g., IgM mesangial proliferative glomerulonephritis and glomerulonephritis caused by autoimmune disorders; such as Goodpasture's syndrome), and nephrocalcinosis.

The recitation of “Digestive” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”) and diseases or disorders of the gastrointestinal system (e.g., as described below under “Gastrointestinal Disorders”.

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Digestive” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: ulcerative colitis, appendicitis, Crohn's disease, hepatitis, hepatic encephalopathy, portal hypertension, cholelithiasis, cancer of the digestive system (e.g., biliary tract cancer, stomach cancer, colon cancer, gastric cancer, pancreatic cancer, cancer of the bile duct, tumors of the colon (e.g., polyps or cancers), and cirrhosis), pancreatitis, ulcerative disease, pyloric stenosis, gastroenteritis, gastritis, gastric atropy, benign tumors of the duodenum, distension, irritable bowel syndrome, malabsorption, congenital disorders of the small intestine, bacterial and parasitic infection, megacolon, Hirschsprung's disease, aganglionic megacolon, acquired megacolon, colitis, anorectal disorders (e.g., anal fistulas, hemorrhoids), congenital disorders of the liver (e.g., Wilson's disease, hemochromatosis, cystic fibrosis, biliary atresia, and alpha1-antitrypsin deficiency), portal hypertension, cholelithiasis, and jaundice.

The recitation of “Connective/Epithelial” in the “Preferred Indication” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under “Hyperproliferative Disorders”), cellular and genetic abnormalities (e.g., as described below under “Diseases at the Cellular Level”), angiogenesis (e.g., as described below under “Anti-Angiogenesis Activity”), and or to promote or inhibit regeneration (e.g., as described below under “Regeneration”), and wound healing (e.g., as described below under “Wound Healing and Epithelial Cell Proliferation”).

In specific embodiments, a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) having a “Connective/Epithelial” recitation in the “Preferred Indication” column of Table 1D, may be used for example, to diagnose, treat, prevent, and/or ameliorate a disease or disorder selected from the group consisting of: connective tissue metaplasia, mixed connective tissue disease, focal epithelial hyperplasia, epithelial metaplasia, mucoepithelial dysplasia, graft v. host disease, polymyositis, cystic hyperplasia, cerebral dysplasia, tissue hypertrophy, Alzheimer's disease, lymphoproliferative disorder, Waldenstron's macroglobulinemia, Crohn's disease, pernicious anemia, idiopathic Addison's disease, glomerulonephritis, bullous pemphigoid, Sjogren's syndrome, diabetes mellitus, cystic fibrosis, osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, osteoporosis, osteocarthritis, periodontal disease, wound healing, relapsing polychondritis, vasculitis, polyarteritis nodosa, Wegener's granulomatosis, cellulitis, rheumatoid arthritis, psoriatic arthritis, discoid lupus erythematosus, systemic lupus erythematosus, scleroderma, CREST syndrome, Sjogren's syndrome, polymyositis, dermatomyositis, mixed connective tissue disease, relapsing polychondritis, vasculitis, Henoch-Schonlein syndrome, erythema nodosum, polyarteritis nodosa, temporal (giant cell) arteritis, Takayasu's arteritis, Wegener's granulomatosis, Reiter's syndrome, Behcet's syndrome, ankylosing spondylitis, cellulitis, keloids, Ehler Danlos syndrome, Marfan syndrome, pseudoxantoma elasticum, osteogenese imperfecta, chondrodysplasias, epidermolysis bullosa, Alport syndrome, and cutis laxa.

Table 1E also provides information regarding biological activities and preferred therapeutic uses (i.e. see, “Preferred Indications” column) for polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof). Table 1E also provides information regarding assays which may be used to test polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) for the corresponding biological activities. The first column (“Gene No.”) provides the gene number in the application for each clone identifier. The second column (“cDNA ATCC Deposit No:Z”) provides the unique clone identifier for each clone as previously described and indicated in Tables 1A, 1B, 1C, and 1D. The third column (“AA SEQ ID NO:Y”) indicates the Sequence Listing SEQ ID Number for polypeptide sequences encoded by the corresponding cDNA clones (also as indicated in Tables 1A, 1B, and 2). The fourth column (“Biological Activity”) indicates a biological activity corresponding to the indicated polypeptides (or polynucleotides encoding said polypeptides). The fifth column (“Exemplary Activity Assay”) further describes the corresponding biological activity and also provides information pertaining to the various types of assays which may be performed to test, demonstrate, or quantify the corresponding biological activity. The sixth column (“Preferred Indications”) describes particular embodiments of the invention as well as indications (e.g. pathologies, diseases, disorders, abnormalities, etc.) for which polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) may be used in detecting, diagnosing, preventing, and/or treating.

The present invention is further directed to antibody-based therapies which involve administering antibodies of the invention to an animal, preferably a mammal, and most preferably a human, patient for treating one or more of the disclosed diseases, disorders, or conditions. Therapeutic compounds of the invention include, but are not limited to, antibodies of the invention (including fragments, analogs and derivatives thereof as described herein) and nucleic acids encoding antibodies of the invention (including fragments, analogs and derivatives thereof and anti-idiotypic antibodies as described herein). The antibodies of the invention can be used to treat, inhibit or prevent diseases, disorders or conditions associated with aberrant expression and/or activity of a polypeptide of the invention, including, but not limited to, any one or more of the diseases, disorders, or conditions described herein. The treatment and/or prevention of diseases, disorders, or conditions associated with aberrant expression and/or activity of a polypeptide of the invention includes, but is not limited to, alleviating symptoms associated with those diseases, disorders or conditions. Antibodies of the invention may be provided in pharmaceutically acceptable compositions as known in the art or as described herein.

In a specific and preferred embodiment, the present invention is directed to antibody-based therapies which involve administering antibodies of the invention to an animal, preferably a mammal, and most preferably a human, patient for treating one or more diseases, disorders, or conditions, including but not limited to: neural disorders, immune system disorders, muscular disorders, reproductive disorders, gastrointestinal disorders, pulmonary disorders, cardiovascular disorders, renal disorders, proliferative disorders, and/or cancerous diseases and conditions, and/or as described elsewhere herein. Therapeutic compounds of the invention include, but are not limited to, antibodies of the invention (e.g., antibodies directed to the full length protein expressed on the cell surface of a mammalian cell; antibodies directed to an epitope of a polypeptide of the invention (such as, for example, a predicted linear epitope shown in column 7 of Table 1B.1; or a conformational epitope, including fragments, analogs and derivatives thereof as described herein) and nucleic acids encoding antibodies of the invention (including fragments, analogs and derivatives thereof and anti-idiotypic antibodies as described herein). The antibodies of the invention can be used to treat, inhibit or prevent diseases, disorders or conditions associated with aberrant expression and/or activity of a polypeptide of the invention, including, but not limited to, any one or more of the diseases, disorders, or conditions described herein. The treatment and/or prevention of diseases, disorders, or conditions associated with aberrant expression and/or activity of a polypeptide of the invention includes, but is not limited to, alleviating symptoms associated with those diseases, disorders or conditions. Antibodies of the invention may be provided in pharmaceutically acceptable compositions as known in the art or as described herein.

A summary of the ways in which the antibodies of the present invention may be used therapeutically includes binding polynucleotides or polypeptides of the present invention locally or systemically in the body or by direct cytotoxicity of the antibody, e.g. as mediated by complement (CDC) or by effector cells (ADCC). Some of these approaches are described in more detail below. Armed with the teachings provided herein, one of ordinary skill in the art will know how to use the antibodies of the present invention for diagnostic, monitoring or therapeutic purposes without undue experimentation.

The antibodies of this invention may be advantageously utilized in combination with other monoclonal or chimeric antibodies, or with lymphokines or hematopoietic growth factors (such as, e.g., IL-2, IL-3 and IL-7), for example, which serve to increase the number or activity of effector cells which interact with the antibodies.

The antibodies of the invention may be administered alone or in combination with other types of treatments (e.g., radiation therapy, chemotherapy, hormonal therapy, immunotherapy and anti-tumor agents). Generally, administration of products of a species origin or species reactivity (in the case of antibodies) that is the same species as that of the patient is preferred. Thus, in a preferred embodiment, human antibodies, fragments derivatives, analogs, or nucleic acids, are administered to a human patient for therapy or prophylaxis.

It is preferred to use high affinity and/or potent in vivo inhibiting and/or neutralizing antibodies against polypeptides or polynucleotides of the present invention, fragments or regions thereof, for both immunoassays directed to and therapy of disorders related to polynucleotides or polypeptides, including fragments thereof, of the present invention. Such antibodies, fragments, or regions, will preferably have an affinity for polynucleotides or polypeptides of the invention, including fragments thereof. Preferred binding affinities include those with a dissociation constant or Kd less than 5×10⁻² M, 10⁻² M, 5×10⁻³ M, 10⁻³ M, 5×10⁻⁴ M, 10⁻⁴ M, 5×10⁻⁵ M, 10⁻⁵ M, 5×10⁻⁶ M, 10⁻⁶ M, 5×10⁻⁷ M, 10⁻⁷ M, 5×10⁻⁸ M, 10⁻⁸ M, 5×10⁻⁹ M, 10⁻⁹ M, 5×10⁻¹⁰ M, 10⁻¹⁰ M, 5×10⁻¹¹ M, 10⁻¹¹ M, 5×10⁻¹² M, 10⁻¹² M, 5×10⁻¹³ M, 10⁻¹³ M, 5×10⁻¹⁴ M, 10¹⁴ M, 5×10⁻¹⁵ M, and 10⁻¹⁵ M.

Gene Therapy

In a specific embodiment, nucleic acids comprising sequences encoding antibodies or functional derivatives thereof, are administered to treat, inhibit or prevent a disease or disorder associated with aberrant expression and/or activity of a polypeptide of the invention, by way of gene therapy. Gene therapy refers to therapy performed by the administration to a subject of an expressed or expressible nucleic acid. In this embodiment of the invention, the nucleic acids produce their encoded protein that mediates a therapeutic effect.

Any of the methods for gene therapy available in the art can be used according to the present invention. Exemplary methods are described below.

For general reviews of the methods of gene therapy, see Goldspiel et al., Clinical Pharmacy 12:488-505 (1993); Wu and Wu, Biotherapy 3:87-95 (1991); Tolstoshev, Ann. Rev. Pharmacol. Toxicol. 32:573-596 (1993); Mulligan, Science 260:926-932 (1993); and Morgan and Anderson, Ann. Rev. Biochem. 62:191-217 (1993); May, TIBTECH 11(5):155-215 (1993). Methods commonly known in the art of recombinant DNA technology which can be used are described in Ausubel et al. (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993); and Kriegler, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990).

In a preferred embodiment, the compound comprises nucleic acid sequences encoding an antibody, said nucleic acid sequences being part of expression vectors that express the antibody or fragments or chimeric proteins or heavy or light chains thereof in a suitable host. In particular, such nucleic acid sequences have promoters operably linked to the antibody coding region, said promoter being inducible or constitutive, and, optionally, tissue-specific. In another particular embodiment, nucleic acid molecules are used in which the antibody coding sequences and any other desired sequences are flanked by regions that promote homologous recombination at a desired site in the genome, thus providing for intrachromosomal expression of the antibody encoding nucleic acids (Koller and Smithies, Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); Zijlstra et al., Nature 342:435-438 (1989). In specific embodiments, the expressed antibody molecule is a single chain antibody; alternatively, the nucleic acid sequences include sequences encoding both the heavy and light chains, or fragments thereof, of the antibody.

Delivery of the nucleic acids into a patient may be either direct, in which case the patient is directly exposed to the nucleic acid or nucleic acid-carrying vectors, or indirect, in which case, cells are first transformed with the nucleic acids in vitro, then transplanted into the patient. These two approaches are known, respectively, as in vivo or ex vivo gene therapy.

In a specific embodiment, the nucleic acid sequences are directly administered in vivo, where it is expressed to produce the encoded product. This can be accomplished by any of numerous methods known in the art, e.g., by constructing them as part of an appropriate nucleic acid expression vector and administering it so that they become intracellular, e.g., by infection using defective or attenuated retrovirals or other viral vectors (see U.S. Pat. No. 4,980,286), or by direct injection of naked DNA, or by use of microparticle bombardment (e.g., a gene gun; Biolistic, Dupont), or coating with lipids or cell-surface receptors or transfecting agents, encapsulation in liposomes, microparticles, or microcapsules, or by administering them in linkage to a peptide which is known to enter the nucleus, by administering it in linkage to a ligand subject to receptor-mediated endocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-4432 (1987)) (which can be used to target cell types specifically expressing the receptors), etc. In another embodiment, nucleic acid-ligand complexes can be formed in which the ligand comprises a fusogenic viral peptide to disrupt endosomes, allowing the nucleic acid to avoid lysosomal degradation. In yet another embodiment, the nucleic acid can be targeted in vivo for cell specific uptake and expression, by targeting a specific receptor (see, e.g., PCT Publications WO 92/06180; WO 92/22635; WO92/20316; WO93/14188, WO 93/20221). Alternatively, the nucleic acid can be introduced intracellularly and incorporated within host cell DNA for expression, by homologous recombination (Koller and Smithies, Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); Zijistra et al., Nature 342:435-438 (1989)).

In a specific embodiment, viral vectors that contains nucleic acid sequences encoding an antibody of the invention are used. For example, a retroviral vector can be used (see Miller et al., Meth. Enzymol. 217:581-599 (1993)). These retroviral vectors contain the components necessary for the correct packaging of the viral genome and integration into the host cell DNA. The nucleic acid sequences encoding the antibody to be used in gene therapy are cloned into one or more vectors, which facilitates delivery of the gene into a patient. More detail about retroviral vectors can be found in Boesen et al., Biotherapy 6:291-302 (1994), which describes the use of a retroviral vector to deliver the mdr1 gene to hematopoietic stem cells in order to make the stem cells more resistant to chemotherapy. Other references illustrating the use of retroviral vectors in gene therapy are: Clowes et al., J. Clin. Invest. 93:644-651 (1994); Kiem et al., Blood 83:1467-1473 (1994); Salmons and Gunzberg, Human Gene Therapy 4:129-141 (1993); and Grossman and Wilson, Curr. Opin. in Genetics and Devel. 3:110-114 (1993).

Adenoviruses are other viral vectors that can be used in gene therapy. Adenoviruses are especially attractive vehicles for delivering genes to respiratory epithelia. Adenoviruses naturally infect respiratory epithelia where they cause a mild disease. Other targets for adenovirus-based delivery systems are liver, the central nervous system, endothelial cells, and muscle. Adenoviruses have the advantage of being capable of infecting non-dividing cells. Kozarsky and Wilson, Current Opinion in Genetics and Development 3:499-503 (1993) present a review of adenovirus-based gene therapy. Bout et al., Human Gene Therapy 5:3-10 (1994) demonstrated the use of adenovirus vectors to transfer genes to the respiratory epithelia of rhesus monkeys. Other instances of the use of adenoviruses in gene therapy can be found in Rosenfeld et al., Science 252:431-434 (1991); Rosenfeld et al., Cell 68:143-155 (1992); Mastrangeli et al., J. Clin. Invest. 91:225-234 (1993); PCT Publication WO94/12649; and Wang, et al., Gene Therapy 2:775-783 (1995). In a preferred embodiment, adenovirus vectors are used.

Adeno-associated virus (AAV) has also been proposed for use in gene therapy (Walsh et al., Proc. Soc. Exp. Biol. Med. 204:289-300 (1993); U.S. Pat. No. 5,436,146).

Another approach to gene therapy involves transferring a gene to cells in tissue culture by such methods as electroporation, lipofection, calcium phosphate mediated transfection, or viral infection. Usually, the method of transfer includes the transfer of a selectable marker to the cells. The cells are then placed under selection to isolate those cells that have taken up and are expressing the transferred gene. Those cells are then delivered to a patient.

In this embodiment, the nucleic acid is introduced into a cell prior to administration in vivo of the resulting recombinant cell. Such introduction can be carried out by any method known in the art, including but not limited to transfection, electroporation, microinjection, infection with a viral or bacteriophage vector containing the nucleic acid sequences, cell fusion, chromosome-mediated gene transfer, microcell-mediated gene transfer, spheroplast fusion, etc. Numerous techniques are known in the art for the introduction of foreign genes into cells (see, e.g., Loeffler and Behr, Meth. Enzymol. 217:599-618 (1993); Cohen et al., Meth. Enzymol. 217:618-644 (1993); Cline, Pharmac. Ther. 29:69-92m (1985) and may be used in accordance with the present invention, provided that the necessary developmental and physiological functions of the recipient cells are not disrupted. The technique should provide for the stable transfer of the nucleic acid to the cell, so that the nucleic acid is expressible by the cell and preferably heritable and expressible by its cell progeny.

The resulting recombinant cells can be delivered to a patient by various methods known in the art. Recombinant blood cells (e.g., hematopoietic stem or progenitor cells) are preferably administered intravenously. The amount of cells envisioned for use depends on the desired effect, patient state, etc., and can be determined by one skilled in the art.

Cells into which a nucleic acid can be introduced for purposes of gene therapy encompass any desired, available cell type, and include but are not limited to epithelial cells, endothelial cells, keratinocytes, fibroblasts, muscle cells, hepatocytes; blood cells such as T lymphocytes, B lymphocytes, monocytes, macrophages, neutrophils, eosinophils, megakaryocytes, granulocytes; various stem or progenitor cells, in particular hematopoietic stem or progenitor cells, e.g., as obtained from bone marrow, umbilical cord blood, peripheral blood, fetal liver, etc.

In a preferred embodiment, the cell used for gene therapy is autologous to the patient.

In an embodiment in which recombinant cells are used in gene therapy, nucleic acid sequences encoding an antibody are introduced into the cells such that they are expressible by the cells or their progeny, and the recombinant cells are then administered in vivo for therapeutic effect. In a specific embodiment, stem or progenitor cells are used. Any stem and/or progenitor cells which can be isolated and maintained in vitro can potentially be used in accordance with this embodiment of the present invention (see e.g. PCT Publication WO 94/08598; Stemple and Anderson, Cell 71:973-985 (1992); Rheinwald, Meth. Cell Bio. 21A:229 (1980); and Pittelkow and Scott, Mayo Clinic Proc. 61:771 (1986)).

In a specific embodiment, the nucleic acid to be introduced for purposes of gene therapy comprises an inducible promoter operably linked to the coding region, such that expression of the nucleic acid is controllable by the presence or absence of an appropriate inducer of transcription.

Demonstration of Therapeutic or Prophylactic Activity

The compounds or pharmaceutical compositions of the invention are preferably tested in vitro, and then in vivo for the desired therapeutic or prophylactic activity, prior to use in humans. For example, in vitro assays to demonstrate the therapeutic or prophylactic utility of a compound or pharmaceutical composition include, the effect of a compound on a cell line or a patient tissue sample. The effect of the compound or composition on the cell line and/or tissue sample can be determined utilizing techniques known to those of skill in the art including, but not limited to, rosette formation assays and cell lysis assays. In accordance with the invention, in vitro assays which can be used to determine whether administration of a specific compound is indicated, include in vitro cell culture assays in which a patient tissue sample is grown in culture, and exposed to or otherwise administered a compound, and the effect of such compound upon the tissue sample is observed.

Therapeutic/Prophylactic Administration and Composition

The invention provides methods of treatment, inhibition and prophylaxis by administration to a subject of an effective amount of a compound or pharmaceutical composition of the invention, preferably a polypeptide or antibody of the invention. In a preferred embodiment, the compound is substantially purified (e.g., substantially free from substances that limit its effect or produce undesired side-effects). The subject is preferably an animal, including but not limited to animals such as cows, pigs, horses, chickens, cats, dogs, etc., and is preferably a mammal, and most preferably human.

Formulations and methods of administration that can be employed when the compound comprises a nucleic acid or an immunoglobulin are described above; additional appropriate formulations and routes of administration can be selected from among those described herein below.

Various delivery systems are known and can be used to administer a compound of the invention, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the compound, receptor-mediated endocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-4432 (1987)), construction of a nucleic acid as part of a retroviral or other vector, etc. Methods of introduction include but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compounds or compositions may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration can be systemic or local. In addition, it may be desirable to introduce the pharmaceutical compounds or compositions of the invention into the central nervous system by any suitable route, including intraventricular and intrathecal injection; intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir, such as an Ommaya reservoir. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent.

In a specific embodiment, it may be desirable to administer the pharmaceutical compounds or compositions of the invention locally to the area in need of treatment; this may be achieved by, for example, and not by way of limitation, local infusion during surgery, topical application, e.g., in conjunction with a wound dressing after surgery, by injection, by means of a catheter, by means of a suppository, or by means of an implant, said implant being of a porous, non-porous, or gelatinous material, including membranes, such as sialastic membranes, or fibers. Preferably, when administering a protein, including an antibody, of the invention, care must be taken to use materials to which the protein does not absorb.

In another embodiment, the compound or composition can be delivered in a vesicle, in particular a liposome (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid.)

In yet another embodiment, the compound or composition can be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989)). In another embodiment, polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, J., Macromol. Sci. Rev. Macromol. Chem. 23:61 (1983); see also Levy et al., Science 228:190 (1985); During et al., Ann. Neurol. 25:351 (1989); Howard et al., J. Neurosurg. 71:105 (1989)). In yet another embodiment, a controlled release system can be placed in proximity of the therapeutic target, e.g., the brain, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)).

Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990)).

In a specific embodiment where the compound of the invention is a nucleic acid encoding a protein, the nucleic acid can be administered in vivo to promote expression of its encoded protein, by constructing it as part of an appropriate nucleic acid expression vector and administering it so that it becomes intracellular, e.g., by use of a retroviral vector (see U.S. Pat. No. 4,980,286), or by direct injection, or by use of microparticle bombardment (e.g., a gene gun; Biolistic, Dupont), or coating with lipids or cell-surface receptors or transfecting agents, or by administering it in linkage to a homeobox-like peptide which is known to enter the nucleus (see e.g., Joliot et al., Proc. Natl. Acad. Sci. USA 88:1864-1868 (1991)), etc. Alternatively, a nucleic acid can be introduced intracellularly and incorporated within host cell DNA for expression, by homologous recombination.

The present invention also provides pharmaceutical compositions. Such compositions comprise a therapeutically effective amount of a compound, and a pharmaceutically acceptable carrier. In a specific embodiment, the term “pharmaceutically acceptable” means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. The term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in “Remington's Pharmaceutical Sciences” by E. W. Martin. Such compositions will contain a therapeutically effective amount of the compound, preferably in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

In a preferred embodiment, the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to human beings. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.

The compounds of the invention can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

The amount of the compound of the invention which will be effective in the treatment, inhibition and prevention of a disease or disorder associated with aberrant expression and/or activity of a polypeptide of the invention can be determined by standard clinical techniques. In addition, in vitro assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness of the disease or disorder, and should be decided according to the judgment of the practitioner and each patient's circumstances. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

For antibodies, the dosage administered to a patient is typically 0.1 mg/kg to 100 mg/kg of the patient's body weight. Preferably, the dosage administered to a patient is between 0.1 mg/kg and 20 mg/kg of the patient's body weight, more preferably 1 mg/kg to 10 mg/kg of the patient's body weight. Generally, human antibodies have a longer half-life within the human body than antibodies from other species due to the immune response to the foreign polypeptides. Thus, lower dosages of human antibodies and less frequent administration is often possible. Further, the dosage and frequency of administration of antibodies of the invention may be reduced by enhancing uptake and tissue penetration (e.g., into the brain) of the antibodies by modifications such as, for example, lipidation.

The invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.

Diagnosis and Imaging

Labeled antibodies, and derivatives and analogs thereof, which specifically bind to a polypeptide of interest can be used for diagnostic purposes to detect, diagnose, or monitor diseases, disorders, and/or conditions associated with the aberrant expression and/or activity of a polypeptide of the invention. The invention provides for the detection of aberrant expression of a polypeptide of interest, comprising (a) assaying the expression of the polypeptide of interest in cells or body fluid of an individual using one or more antibodies specific to the polypeptide interest and (b) comparing the level of gene expression with a standard gene expression level, whereby an increase or decrease in the assayed polypeptide gene expression level compared to the standard expression level is indicative of aberrant expression.

The invention provides a diagnostic assay for diagnosing a disorder, comprising (a) assaying the expression of the polypeptide of interest in cells or body fluid of an individual using one or more antibodies specific to the polypeptide interest and (b) comparing the level of gene expression with a standard gene expression level, whereby an increase or decrease in the assayed polypeptide gene expression level compared to the standard expression level is indicative of a particular disorder. With respect to cancer, the presence of a relatively high amount of transcript in biopsied tissue from an individual may indicate a predisposition for the development of the disease, or may provide a means for detecting the disease prior to the appearance of actual clinical symptoms. A more definitive diagnosis of this type may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development or further progression of the cancer.

Antibodies of the invention can be used to assay protein levels in a biological sample using classical immunohistological methods known to those of skill in the art (e.g., see Jalkanen et al., J. Cell. Biol. 101:976-985 (1985); Jalkanen et al., J. Cell. Biol. 105:3087-3096 (1987)). Other antibody-based methods useful for detecting protein gene expression include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA). Suitable antibody assay labels are known in the art and include enzyme labels, such as, glucose oxidase; radioisotopes, such as iodine (125I, 121I), carbon (14C), sulfur (35S), tritium (3H), indium (112In), and technetium (99Tc); luminescent labels, such as luminol; and fluorescent labels, such as fluorescein and rhodamine, and biotin.

One facet of the invention is the detection and diagnosis of a disease or disorder associated with aberrant expression of a polypeptide of interest in an animal, preferably a mammal and most preferably a human. In one embodiment, diagnosis comprises: a) administering (for example, parenterally, subcutaneously, or intraperitoneally) to a subject an effective amount of a labeled molecule which specifically binds to the polypeptide of interest; b) waiting for a time interval following the administering for permitting the labeled molecule to preferentially concentrate at sites in the subject where the polypeptide is expressed (and for unbound labeled molecule to be cleared to background level); c) determining background level; and d) detecting the labeled molecule in the subject, such that detection of labeled molecule above the background level indicates that the subject has a particular disease or disorder associated with aberrant expression of the polypeptide of interest. Background level can be determined by various methods including, comparing the amount of labeled molecule detected to a standard value previously determined for a particular system.

It will be understood in the art that the size of the subject and the imaging system used will determine the quantity of imaging moiety needed to produce diagnostic images. In the case of a radioisotope moiety, for a human subject, the quantity of radioactivity injected will normally range from about 5 to 20 millicuries of 99mTc. The labeled antibody or antibody fragment will then preferentially accumulate at the location of cells which contain the specific protein. In vivo tumor imaging is described in S. W. Burchiel et al., “Immunopharmacokinetics of Radiolabeled Antibodies and Their Fragments.” (Chapter 13 in Tumor Imaging: The Radiochemical Detection of Cancer, S. W. Burchiel and B. A. Rhodes, eds., Masson Publishing Inc. (1982)).

Depending on several variables, including the type of label used and the mode of administration, the time interval following the administration for permitting the labeled molecule to preferentially concentrate at sites in the subject and for unbound labeled molecule to be cleared to background level is 6 to 48 hours or 6 to 24 hours or 6 to 12 hours. In another embodiment the time interval following administration is 5 to 20 days or 5 to 10 days.

In an embodiment, monitoring of the disease or disorder is carried out by repeating the method for diagnosing the disease or disease, for example, one month after initial diagnosis, six months after initial diagnosis, one year after initial diagnosis, etc.

Presence of the labeled molecule can be detected in the patient using methods known in the art for in vivo scanning. These methods depend upon the type of label used. Skilled artisans will be able to determine the appropriate method for detecting a particular label. Methods and devices that may be used in the diagnostic methods of the invention include, but are not limited to, computed tomography (CT), whole body scan such as position emission tomography (PET), magnetic resonance imaging (MRI), and sonography.

In a specific embodiment, the molecule is labeled with a radioisotope and is detected in the patient using a radiation responsive surgical instrument (Thurston et al., U.S. Pat. No. 5,441,050). In another embodiment, the molecule is labeled with a fluorescent compound and is detected in the patient using a fluorescence responsive scanning instrument. In another embodiment, the molecule is labeled with a positron emitting metal and is detected in the patent using positron emission-tomography. In yet another embodiment, the molecule is labeled with a paramagnetic label and is detected in a patient using magnetic resonance imaging (MRI).

Kits

The present invention provides kits that can be used in the above methods. In one embodiment, a kit comprises an antibody of the invention, preferably a purified antibody, in one or more containers. In a specific embodiment, the kits of the present invention contain a substantially isolated polypeptide comprising an epitope which is specifically immunoreactive with an antibody included in the kit. Preferably, the kits of the present invention further comprise a control antibody which does not react with the polypeptide of interest. In another specific embodiment, the kits of the present invention contain a means for detecting the binding of an antibody to a polypeptide of interest (e.g., the antibody may be conjugated to a detectable substrate such as a fluorescent compound, an enzymatic substrate, a radioactive compound or a luminescent compound, or a second antibody which recognizes the first antibody may be conjugated to a detectable substrate).

In another specific embodiment of the present invention, the kit is a diagnostic kit for use in screening serum containing antibodies specific against proliferative and/or cancerous polynucleotides and polypeptides. Such a kit may include a control antibody that does not react with the polypeptide of interest. Such a kit may include a substantially isolated polypeptide antigen comprising an epitope which is specifically immunoreactive with at least one anti-polypeptide antigen antibody. Further, such a kit includes means for detecting the binding of said antibody to the antigen (e.g., the antibody may be conjugated to a fluorescent compound such as fluorescein or rhodamine which can be detected by flow cytometry). In specific embodiments, the kit may include a recombinantly produced or chemically synthesized polypeptide antigen. The polypeptide antigen of the kit may also be attached to a solid support.

In a more specific embodiment the detecting means of the above-described kit includes a solid support to which said polypeptide antigen is attached. Such a kit may also include a non-attached reporter-labeled anti-human antibody. In this embodiment, binding of the antibody to the polypeptide antigen can be detected by binding of the said reporter-labeled antibody.

In an additional embodiment, the invention includes a diagnostic kit for use in screening serum containing antigens of the polypeptide of the invention. The diagnostic kit includes a substantially isolated antibody specifically immunoreactive with polypeptide or polynucleotide antigens, and means for detecting the binding of the polynucleotide or polypeptide antigen to the antibody. In one embodiment, the antibody is attached to a solid support. In a specific embodiment, the antibody may be a monoclonal antibody. The detecting means of the kit may include a second, labeled monoclonal antibody. Alternatively, or in addition, the detecting means may include a labeled, competing antigen.

In one diagnostic configuration, test serum is reacted with a solid phase reagent having a surface-bound antigen obtained by the methods of the present invention. After binding with specific antigen antibody to the reagent and removing unbound serum components by washing, the reagent is reacted with reporter-labeled anti-human antibody to bind reporter to the reagent in proportion to the amount of bound anti-antigen antibody on the solid support. The reagent is again washed to remove unbound labeled antibody, and the amount of reporter associated with the reagent is determined. Typically, the reporter is an enzyme which is detected by incubating the solid phase in the presence of a suitable fluorometric, luminescent or colorimetric substrate (Sigma, St. Louis, Mo.).

The solid surface reagent in the above assay is prepared by known techniques for attaching protein material to solid support material, such as polymeric beads, dip sticks, 96-well plate or filter material. These attachment methods generally include non-specific adsorption of the protein to the support or covalent attachment of the protein, typically through a free amine group, to a chemically reactive group on the solid support, such as an activated carboxyl, hydroxyl, or aldehyde group. Alternatively, streptavidin coated plates can be used in conjunction with biotinylated antigen(s).

Thus, the invention provides an assay system or kit for carrying out this diagnostic method. The kit generally includes a support with surface-bound recombinant antigens, and a reporter-labeled anti-human antibody for detecting surface-bound anti-antigen antibody.

Uses of the Polynucleotides

Each of the polynucleotides identified herein can be used in numerous ways as reagents. The following description should be considered exemplary and utilizes known techniques.

The polynucleotides of the present invention are useful for chromosome identification. There exists an ongoing need to identify new chromosome markers, since few chromosome marking reagents, based on actual sequence data (repeat polymorphisms), are presently available. Each sequence is specifically targeted to and can hybridize with a particular location on an individual human chromosome, thus each polynucleotide of the present invention can routinely be used as a chromosome marker using techniques known in the art. Table 1B.1, column 8 provides the chromosome location of some of the polynucleotides of the invention.

Briefly, sequences can be mapped to chromosomes by preparing PCR primers (preferably at least 15 bp (e.g., 15-25 bp) from the sequences shown in SEQ ID NO:X. Primers can optionally be selected using computer analysis so that primers do not span more than one predicted exon in the genomic DNA. These primers are then used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those hybrids containing the human gene corresponding to SEQ ID NO:X will yield an amplified fragment.

Similarly, somatic hybrids provide a rapid method of PCR mapping the polynucleotides to particular chromosomes. Three or more clones can be assigned per day using a single thermal cycler. Moreover, sublocalization of the polynucleotides can be achieved with panels of specific chromosome fragments. Other gene mapping strategies that can be used include in situ hybridization, prescreening with labeled flow-sorted chromosomes, preselection by hybridization to construct chromosome specific-cDNA libraries, and computer mapping techniques (See, e.g., Shuler, Trends Biotechnol 16:456-459 (1998) which is hereby incorporated by reference in its entirety).

Precise chromosomal location of the polynucleotides can also be achieved using fluorescence in situ hybridization (FISH) of a metaphase chromosomal spread. This technique uses polynucleotides as short as 500 or 600 bases; however, polynucleotides 2,000 bp are preferred. For a review of this technique, see Verma et al., “Human Chromosomes: a Manual of Basic Techniques,” Pergamon Press, New York (1988).

For chromosome mapping, the polynucleotides can be used individually (to mark a single chromosome or a single site on that chromosome) or in panels (for marking multiple sites and/or multiple chromosomes).

Thus, the present invention also provides a method for chromosomal localization which involves (a) preparing PCR primers from the polynucleotide sequences in Table 1B and/or Table 2 and SEQ ID NO:X and (b) screening somatic cell hybrids containing individual chromosomes.

The polynucleotides of the present invention would likewise be useful for radiation hybrid mapping, HAPPY mapping, and long range restriction mapping. For a review of these techniques and others known in the art, see, e.g. Dear, “Genome Mapping: A Practical Approach,” IRL Press at Oxford University Press, London (1997); Aydin, J. Mol. Med. 77:691-694 (1999); Hacia et al., Mol. Psychiatry 3:483-492 (1998); Herrick et al., Chromosome Res. 7:409-423 (1999); Hamilton et al., Methods Cell Biol. 62:265-280 (2000); and/or Ott, J. Hered. 90:68-70 (1999) each of which is hereby incorporated by reference in its entirety.

Once a polynucleotide has been mapped to a precise chromosomal location, the physical position of the polynucleotide can be used in linkage analysis. Linkage analysis establishes coinheritance between a chromosomal location and presentation of a particular disease. (Disease mapping data are found, for example, in V. McKusick, Mendelian Inheritance in Man (available on line through Johns Hopkins University Welch Medical Library)). Column 9 of Table 1B.1 provides an OMIM reference identification number of diseases associated with the cytologic band disclosed in column 8 of Table 1B.1, as determined using techniques described herein and by reference to Table 5. Assuming 1 megabase mapping resolution and one gene per 20 kb, a cDNA precisely localized to a chromosomal region associated with the disease could be one of 50-500 potential causative genes.

Thus, once coinheritance is established, differences in a polynucleotide of the invention and the corresponding gene between affected and unaffected individuals can be examined. First, visible structural alterations in the chromosomes, such as deletions or translocations, are examined in chromosome spreads or by PCR. If no structural alterations exist, the presence of point mutations are ascertained. Mutations observed in some or all affected individuals, but not in normal individuals, indicates that the mutation may cause the disease. However, complete sequencing of the polypeptide and the corresponding gene from several normal individuals is required to distinguish the mutation from a polymorphism. If a new polymorphism is identified, this polymorphic polypeptide can be used for further linkage analysis.

Furthermore, increased or decreased expression of the gene in affected individuals as compared to unaffected individuals can be assessed using the polynucleotides of the invention. Any of these alterations (altered expression, chromosomal rearrangement, or mutation) can be used as a diagnostic or prognostic marker. Diagnostic and prognostic methods, kits and reagents encompassed by the present invention are briefly described below and more thoroughly elsewhere herein (see e.g., the sections labeled “Antibodies”, “Diagnostic Assays”, and “Methods for Detecting Diseases”).

Thus, the invention also provides a diagnostic method useful during diagnosis of a disorder, involving measuring the expression level of polynucleotides of the present invention in cells or body fluid from an individual and comparing the measured gene expression level with a standard level of polynucleotide expression level, whereby an increase or decrease in the gene expression level compared to the standard is indicative of a disorder. Additional non-limiting examples of diagnostic methods encompassed by the present invention are more thoroughly described elsewhere herein (see, e.g., Example 12).

In still another embodiment, the invention includes a kit for analyzing samples for the presence of proliferative and/or cancerous polynucleotides derived from a test subject. In a general embodiment, the kit includes at least one polynucleotide probe containing a nucleotide sequence that will specifically hybridize with a polynucleotide of the invention and a suitable container. In a specific embodiment, the kit includes two polynucleotide probes defining an internal region of the polynucleotide of the invention, where each probe has one strand containing a 31′mer-end internal to the region. In a further embodiment, the probes may be useful as primers for polymerase chain reaction amplification.

Where a diagnosis of a related disorder, including, for example, diagnosis of a tumor, has already been made according to conventional methods, the present invention is useful as a prognostic indicator, whereby patients exhibiting enhanced or depressed polynucleotide of the invention expression will experience a worse clinical outcome relative to patients expressing the gene at a level nearer the standard level.

By “measuring the expression level of polynucleotides of the invention” is intended qualitatively or quantitatively measuring or estimating the level of the polypeptide of the invention or the level of the mRNA encoding the polypeptide of the invention in a first biological sample either directly (e.g., by determining or estimating absolute protein level or mRNA level) or relatively (e.g., by comparing to the polypeptide level or mRNA level in a second biological sample). Preferably, the polypeptide level or mRNA level in the first biological sample is measured or estimated and compared to a standard polypeptide level or mRNA level, the standard being taken from a second biological sample obtained from an individual not having the related disorder or being determined by averaging levels from a population of individuals not having a related disorder. As will be appreciated in the art, once a standard polypeptide level or mRNA level is known, it can be used repeatedly as a standard for comparison.

By “biological sample” is intended any biological sample obtained from an individual, body fluid, cell line, tissue culture, or other source which contains polypeptide of the present invention or the corresponding mRNA. As indicated, biological samples include body fluids (such as semen, lymph, vaginal pool, sera, plasma, urine, synovial fluid and spinal fluid) which contain the polypeptide of the present invention, and tissue sources found to express the polypeptide of the present invention. Methods for obtaining tissue biopsies and body fluids from mammals are well known in the art. Where the biological sample is to include mRNA, a tissue biopsy is the preferred source.

The method(s) provided above may preferably be applied in a diagnostic method and/or kits in which polynucleotides and/or polypeptides of the invention are attached to a solid support. In one exemplary method, the support may be a “gene chip” or a “biological chip” as described in U.S. Pat. Nos. 5,837,832, 5,874,219, and 5,856,174. Further, such a gene chip with polynucleotides of the invention attached may be used to identify polymorphisms between the isolated polynucleotide sequences of the invention, with polynucleotides isolated from a test subject. The knowledge of such polymorphisms (i.e. their location, as well as, their existence) would be beneficial in identifying disease loci for many disorders, such as for example, in neural disorders, immune system disorders, muscular disorders, reproductive disorders, gastrointestinal disorders, pulmonary disorders, digestive disorders, metabolic disorders, cardiovascular disorders, renal disorders, proliferative disorders, and/or cancerous diseases and conditions. Such a method is described in U.S. Pat. Nos. 5,858,659 and 5,856,104. The US patents referenced supra are hereby incorporated by reference in their entirety herein.

The present invention encompasses polynucleotides of the present invention that are chemically synthesized, or reproduced as peptide nucleic acids (PNA), or according to other methods known in the art. The use of PNAs would serve as the preferred form if the polynucleotides of the invention are incorporated onto a solid support, or gene chip. For the purposes of the present invention, a peptide nucleic acid (PNA) is a polyamide type of DNA analog and the monomeric units for adenine, guanine, thymine and cytosine are available commercially (Perceptive Biosystems). Certain components of DNA, such as phosphorus, phosphorus oxides, or deoxyribose derivatives, are not present in PNAs. As disclosed by Nielsen et al., Science 254, 1497 (1991); and Egholm et al., Nature 365, 666 (1993), PNAs bind specifically and tightly to complementary DNA strands and are not degraded by nucleases. In fact, PNA binds more strongly to DNA than DNA itself does. This is probably because there is no electrostatic repulsion between the two strands, and also the polyamide backbone is more flexible. Because of this, PNA/DNA duplexes bind under a wider range of stringency conditions than DNA/DNA duplexes, making it easier to perform multiplex hybridization. Smaller probes can be used than with DNA due to the strong binding. In addition, it is more likely that single base mismatches can be determined with PNA/DNA hybridization because a single mismatch in a PNA/DNA 15-mer lowers the melting point (T.sub.m) by 8°-20° C., vs. 4°-16° C. for the DNA/DNA 15-mer duplex. Also, the absence of charge groups in PNA means that hybridization can be done at low ionic strengths and reduce possible interference by salt during the analysis.

The compounds of the present invention have uses which include, but are not limited to, detecting cancer in mammals. In particular the invention is useful during diagnosis of pathological cell proliferative neoplasias which include, but are not limited to: acute myelogenous leukemias including acute monocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute erythroleukemia, acute megakaryocytic leukemia, and acute undifferentiated leukemia, etc.; and chronic myelogenous leukemias including chronic myelomonocytic leukemia, chronic granulocytic leukemia, etc. Preferred mammals include monkeys, apes, cats, dogs, cows, pigs, horses, rabbits and humans. Particularly preferred are humans.

Pathological cell proliferative disorders are often associated with inappropriate activation of proto-oncogenes. (Gelmann, E. P. et al., “The Etiology of Acute Leukemia: Molecular Genetics and Viral Oncology,” in Neoplastic Diseases of the Blood, Vol 1., Wiernik, P. H. et al. eds., 161-182 (1985)). Neoplasias are now believed to result from the qualitative alteration of a normal cellular gene product, or from the quantitative modification of gene expression by insertion into the chromosome of a viral sequence, by chromosomal translocation of a gene to a more actively transcribed region, or by some other mechanism. (Gelmann et al., supra) It is likely that mutated or altered expression of specific genes is involved in the pathogenesis of some leukemias, among other tissues and cell types. (Gelmann et al., supra) Indeed, the human counterparts of the oncogenes involved in some animal neoplasias have been amplified or translocated in some cases of human leukemia and carcinoma. (Gelmann et al., supra)

For example, c-myc expression is highly amplified in the non-lymphocytic leukemia cell line HL-60. When HL-60 cells are chemically induced to stop proliferation, the level of c-myc is found to be downregulated. (International Publication Number WO 91/15580). However, it has been shown that exposure of HL-60 cells to a DNA construct that is complementary to the 5′ end of c-myc or c-myb blocks translation of the corresponding mRNAs which downregulates expression of the c-myc or c-myb proteins and causes arrest of cell proliferation and differentiation of the treated cells. (International Publication Number WO 91/15580; Wickstrom et al., Proc. Natl. Acad. Sci. 85:1028 (1988); Anfossi et al., Proc. Natl. Acad. Sci. 86:3379 (1989)). However, the skilled artisan would appreciate the present invention's usefulness is not be limited to treatment, prevention, and/or prognosis of proliferative disorders of cells and tissues of hematopoietic origin, in light of the numerous cells and cell types of varying origins which are known to exhibit proliferative phenotypes.

In addition to the foregoing, a polynucleotide of the present invention can be used to control gene expression through triple helix formation or through antisense DNA or RNA. Antisense techniques are discussed, for example, in Okano, J. Neurochem. 56: 560 (1991); “Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, Fla. (1988). Triple helix formation is discussed in, for instance Lee et al., Nucleic Acids Research 6: 3073 (1979); Cooney et al., Science 241: 456 (1988); and Dervan et al., Science 251: 1360 (1991). Both methods rely on binding of the polynucleotide to a complementary DNA or RNA. For these techniques, preferred polynucleotides are usually oligonucleotides 20 to 40 bases in length and complementary to either the region of the gene involved in transcription (triple helix—see Lee et al., Nucl. Acids Res. 6:3073 (1979); Cooney et al., Science 241:456 (1988); and Dervan et al., Science 251:1360 (1991)) or to the mRNA itself (antisense—Okano, J. Neurochem. 56:560 (1991); Oligodeoxy-nucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, Fla. (1988)). Triple helix formation optimally results in a shut-off of RNA transcription from DNA, while antisense RNA hybridization blocks translation of an mRNA molecule into polypeptide. The oligonucleotide described above can also be delivered to cells such that the antisense RNA or DNA may be expressed in vivo to inhibit production of polypeptide of the present invention antigens. Both techniques are effective in model systems, and the information disclosed herein can be used to design antisense or triple helix polynucleotides in an effort to treat disease, and in particular, for the treatment of proliferative diseases and/or conditions. Non-limiting antisense and triple helix methods encompassed by the present invention are more thoroughly described elsewhere herein (see, e.g., the section labeled “Antisense and Ribozyme (Antagonists)”).

Polynucleotides of the present invention are also useful in gene therapy. One goal of gene therapy is to insert a normal gene into an organism having a defective gene, in an effort to correct the genetic defect. The polynucleotides disclosed in the present invention offer a means of targeting such genetic defects in a highly accurate manner. Another goal is to insert a new gene that was not present in the host genome, thereby producing a new trait in the host cell. Additional non-limiting examples of gene therapy methods encompassed by the present invention are more thoroughly described elsewhere herein (see, e.g., the sections labeled “Gene Therapy Methods”, and Examples 16, 17 and 18).

The polynucleotides are also useful for identifying individuals from minute biological samples. The United States military, for example, is considering the use of restriction fragment length polymorphism (RFLP) for identification of its personnel. In this technique, an individual's genomic DNA is digested with one or more restriction enzymes, and probed on a Southern blot to yield unique bands for identifying personnel. This method does not suffer from the current limitations of “Dog Tags” which can be lost, switched, or stolen, making positive identification difficult. The polynucleotides of the present invention can be used as additional DNA markers for RFLP.

The polynucleotides of the present invention can also be used as an alternative to RFLP, by determining the actual base-by-base DNA sequence of selected portions of an individual's genome. These sequences can be used to prepare PCR primers for amplifying and isolating such selected DNA, which can then be sequenced. Using this technique, individuals can be identified because each individual will have a unique set of DNA sequences. Once an unique ID database is established for an individual, positive identification of that individual, living or dead, can be made from extremely small tissue samples.

Forensic biology also benefits from using DNA-based identification techniques as disclosed herein. DNA sequences taken from very small biological samples such as tissues, e.g., hair or skin, or body fluids, e.g., blood, saliva, semen, synovial fluid, amniotic fluid, breast milk, lymph, pulmonary sputum or surfactant, urine, fecal matter, etc., can be amplified using PCR. In one prior art technique, gene sequences amplified from polymorphic loci, such as DQa class II HLA gene, are used in forensic biology to identify individuals. (Erlich, H., PCR Technology, Freeman and Co. (1992)). Once these specific polymorphic loci are amplified, they are digested with one or more restriction enzymes, yielding an identifying set of bands on a Southern blot probed with DNA corresponding to the DQa class II HLA gene. Similarly, polynucleotides of the present invention can be used as polymorphic markers for forensic purposes.

There is also a need for reagents capable of identifying the source of a particular tissue. Such need arises, for example, in forensics when presented with tissue of unknown origin. Appropriate reagents can comprise, for example, DNA probes or primers prepared from the sequences of the present invention, specific to tissues, including but not limited to those shown in Table 1B. Panels of such reagents can identify tissue by species and/or by organ type. In a similar fashion, these reagents can be used to screen tissue cultures for contamination. Additional non-limiting examples of such uses are further described herein.

The polynucleotides of the present invention are also useful as hybridization probes for differential identification of the tissue(s) or cell type(s) present in a biological sample. Similarly, polypeptides and antibodies directed to polypeptides of the present invention are useful to provide immunological probes for differential identification of the tissue(s) (e.g., immunohistochemistry assays) or cell type(s) (e.g., immunocytochemistry assays). In addition, for a number of disorders of the above tissues or cells, significantly higher or lower levels of gene expression of the polynucleotides/polypeptides of the present invention may be detected in certain tissues (e.g., tissues expressing polypeptides and/or polynucleotides of the present invention, for example, those disclosed in column 5 of Table 1B.2, and/or cancerous and/or wounded tissues) or bodily fluids (e.g., semen, lymph, vaginal pool, serum, plasma, urine, synovial fluid or spinal fluid) taken from an individual having such a disorder, relative to a “standard” gene expression level, i.e., the expression level in healthy tissue from an individual not having the disorder.

Thus, the invention provides a diagnostic method of a disorder, which involves: (a) assaying gene expression level in cells or body fluid of an individual; (b) comparing the gene expression level with a standard gene expression level, whereby an increase or decrease in the assayed gene expression level compared to the standard expression level is indicative of a disorder.

In the very least, the polynucleotides of the present invention can be used as molecular weight markers on Southern gels, as diagnostic probes for the presence of a specific mRNA in a particular cell type, as a probe to “subtract-out” known sequences in the process of discovering novel polynucleotides, for selecting and making oligomers for attachment to a “gene chip” or other support, to raise anti-DNA antibodies using DNA immunization techniques, and as an antigen to elicit an immune response.

Uses of the Polypeptides

Each of the polypeptides identified herein can be used in numerous ways. The following description should be considered exemplary and utilizes known techniques.

Polypeptides and antibodies directed to polypeptides of the present invention are useful to provide immunological probes for differential identification of the tissue(s) (e.g., immunohistochemistry assays such as, for example, ABC immunoperoxidase (Hsu et al., J. Histochem. Cytochem. 29:577-580 (1981)) or cell type(s) (e.g., immunocytochemistry assays).

Antibodies can be used to assay levels of polypeptides encoded by polynucleotides of the invention in a biological sample using classical immunohistological methods known to those of skill in the art (e.g., see Jalkanen, et al., J. Cell. Biol. 101:976-985 (1985); Jalkanen, et al., J. Cell. Biol. 105:3087-3096 (1987)). Other antibody-based methods useful for detecting protein gene expression include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA). Suitable antibody assay labels are known in the art and include enzyme labels, such as, glucose oxidase; radioisotopes, such as iodine (¹³¹I, ¹²⁵I, ¹²³I, ¹²¹I), carbon (¹⁴C), sulfur (³⁵S), tritium (³H), indium (^(115m)In, ^(113m)In, ¹¹²In, ¹¹¹In), and technetium (⁹⁹Tc, ^(99m)Tc), thallium (²⁰¹Ti), gallium (⁶⁸Ga, ⁶⁷Ga), palladium (¹⁰³Pd), molybdenum (⁹⁹Mo), xenon (¹³³Xe), fluorine (¹⁸F), ¹⁵³Sm, ¹⁷⁷Lu, ¹⁵⁹Gd, ¹⁴⁹Pm, ¹⁴⁰La, ¹⁷⁵Yb, ¹⁶⁶Ho, ⁹⁰Y, ⁴⁷Sc, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁴²Pr, ¹⁰⁵Rh, ⁹⁷Ru; luminescent labels, such as luminol; and fluorescent labels, such as fluorescein and rhodamine, and biotin.

In addition to assaying levels of polypeptide of the present invention in a biological sample, proteins can also be detected in vivo by imaging. Antibody labels or markers for in vivo imaging of protein include those detectable by X-radiography, NMR or ESR. For X-radiography, suitable labels include radioisotopes such as barium or cesium, which emit detectable radiation but are not overtly harmful to the subject. Suitable markers for NMR and ESR include those with a detectable characteristic spin, such as deuterium, which may be incorporated into the antibody by labeling of nutrients for the relevant hybridoma.

A protein-specific antibody or antibody fragment which has been labeled with an appropriate detectable imaging moiety, such as a radioisotope (for example, ¹³¹I, ¹¹²In, ^(99m)Tc, (¹³¹I, ¹²⁵I, ¹²³I, ¹²¹I), carbon (¹⁴C), sulfur (³⁵S), tritium (³H), indium (^(115m)In, ^(113m)In, ¹¹²In, ¹¹¹In), and technetium (⁹⁹Tc, ^(99m)Tc), thallium (²⁰¹Ti), gallium (⁶⁸Ga, ⁶⁷Ga), palladium (¹⁰³Pd), molybdenum (⁹⁹Mo), xenon (¹³³Xe), fluorine (¹⁸F, ¹⁵³Sm, ¹⁷⁷Lu, ¹⁵⁹Gd, ¹⁴⁹Pm, ¹⁴⁰La, ¹⁷⁵Yb, ¹⁶⁶Ho, ⁹⁰Y, ⁴⁷Sc, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁴²Pr, ¹⁰⁵Rh, ⁹⁷Ru), a radio-opaque substance, or a material detectable by nuclear magnetic resonance, is introduced (for example, parenterally, subcutaneously or intraperitoneally) into the mammal to be examined for immune system disorder. It will be understood in the art that the size of the subject and the imaging system used will determine the quantity of imaging moiety needed to produce diagnostic images. In the case of a radioisotope moiety, for a human subject, the quantity of radioactivity injected will normally range from about 5 to 20 millicuries of ^(99m)Tc. The labeled antibody or antibody fragment will then preferentially accumulate at the location of cells which express the polypeptide encoded by a polynucleotide of the invention. In vivo tumor imaging is described in S. W. Burchiel et al., “Immunopharmacokinctics of Radiolabeled Antibodies and Their Fragments” (Chapter 13 in Tumor Imaging: The Radiochemical Detection of Cancer, S. W. Burchiel and B. A. Rhodes, eds., Masson Publishing Inc. (1982)).

In one embodiment, the invention provides a method for the specific delivery of compositions of the invention to cells by administering polypeptides of the invention (e.g., polypeptides encoded by polynucleotides of the invention and/or antibodies) that are associated with heterologous polypeptides or nucleic acids. In one example, the invention provides a method for delivering a therapeutic protein into the targeted cell. In another example, the invention provides a method for delivering a single stranded nucleic acid (e.g., antisense or ribozymes) or double stranded nucleic acid (e.g., DNA that can integrate into the cell's genome or replicate episomally and that can be transcribed) into the targeted cell.

In another embodiment, the invention provides a method for the specific destruction of cells (e.g., the destruction of tumor cells) by administering polypeptides of the invention in association with toxins or cytotoxic prodrugs.

By “toxin” is meant one or more compounds that bind and activate endogenous cytotoxic effector systems, radioisotopes, holotoxins, modified toxins, catalytic subunits of toxins, or any molecules or enzymes not normally present in or on the surface of a cell that under defined conditions cause the cell's death. Toxins that may be used according to the methods of the invention include, but are not limited to, radioisotopes known in the art, compounds such as, for example, antibodies (or complement fixing containing portions thereof) that bind an inherent or induced endogenous cytotoxic effector system, thymidine kinase, endonuclease, RNAse, alpha toxin, ricin, abrin, Pseudomonas exotoxin A, diphtheria toxin, saporin, momordin, gelonin, pokeweed antiviral protein, alpha-sarcin and cholera toxin. “Toxin” also includes a cytostatic or cytocidal agent, a therapeutic agent or a radioactive metal ion, e.g., alpha-emitters such as, for example, ²¹³Bi, or other radioisotopes such as, for example, ¹⁰³Pd, ¹³³Xe, ¹³¹I, ⁶⁸Ge, ⁵⁷Co, ⁶⁵Zn, ⁸⁵Sr, ³²P, ³⁵S, ⁹⁰Y, ¹⁵³Sm, ¹⁵³Gd, ¹⁶⁹Yb, ⁵¹Cr, ⁵⁴Mn, ⁷⁵Se, ¹¹³Sn, ⁹⁰Yttrium, ¹¹⁷Tin, ¹⁸⁶Rhenium, ¹⁶⁶Holmium, and ¹⁸⁸Rhenium; luminescent labels, such as luminol; and fluorescent labels, such as fluorescein and rhodamine, and biotin. In a specific embodiment, the invention provides a method for the specific destruction of cells (e.g., the destruction of tumor cells) by administering polypeptides of the invention or antibodies of the invention in association with the radioisotope ⁹⁰Y. In another specific embodiment, the invention provides a method for the specific destruction of cells (e.g., the destruction of tumor cells) by administering polypeptides of the invention or antibodies of the invention in association with the radioisotope ¹¹¹In. In a further specific embodiment, the invention provides a method for the specific destruction of cells (e.g., the destruction of tumor cells) by administering polypeptides of the invention or antibodies of the invention in association with the radioisotope ¹³¹I.

Techniques known in the art may be applied to label polypeptides of the invention (including antibodies). Such techniques include, but are not limited to, the use of bifunctional conjugating agents (see e.g., U.S. Pat. Nos. 5,756,065; 5,714,631; 5,696,239; 5,652,361; 5,505,931; 5,489,425; 5,435,990; 5,428,139; 5,342,604; 5,274,119; 4,994,560; and 5,808,003; the contents of each of which are hereby incorporated by reference in its entirety).

Thus, the invention provides a diagnostic method of a disorder, which involves (a) assaying the expression level of a polypeptide of the present invention in cells or body fluid of an individual; and (b) comparing the assayed polypeptide expression level with a standard polypeptide expression level, whereby an increase or decrease in the assayed polypeptide expression level compared to the standard expression level is indicative of a disorder. With respect to cancer, the presence of a relatively high amount of transcript in biopsied tissue from an individual may indicate a predisposition for the development of the disease, or may provide a means for detecting the disease prior to the appearance of actual clinical symptoms. A more definitive diagnosis of this type may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development or further progression of the cancer.

Moreover, polypeptides of the present invention can be used to treat or prevent diseases or conditions such as, for example, neural disorders, immune system disorders, muscular disorders, reproductive disorders, gastrointestinal disorders, pulmonary disorders, cardiovascular disorders, renal disorders, proliferative disorders, and/or cancerous diseases and conditions. For example, patients can be administered a polypeptide of the present invention in an effort to replace absent or decreased levels of the polypeptide (e.g., insulin), to supplement absent or decreased levels of a different polypeptide (e.g., hemoglobin S for hemoglobin B, SOD, catalase, DNA repair proteins), to inhibit the activity of a polypeptide (e.g., an oncogene or tumor supressor), to activate the activity of a polypeptide (e.g., by binding to a receptor), to reduce the activity of a membrane bound receptor by competing with it for free ligand (e.g., soluble TNF receptors used in reducing inflammation), or to bring about a desired response (e.g., blood vessel growth inhibition, enhancement of the immune response to proliferative cells or tissues).

Similarly, antibodies directed to a polypeptide of the present invention can also be used to treat disease (as described supra, and elsewhere herein). For example, administration of an antibody directed to a polypeptide of the present invention can bind, and/or neutralize the polypeptide, and/or reduce overproduction of the polypeptide. Similarly, administration of an antibody can activate the polypeptide, such as by binding to a polypeptide bound to a membrane (receptor).

At the very least, the polypeptides of the present invention can be used as molecular weight markers on SDS-PAGE gels or on molecular sieve gel filtration columns using methods well known to those of skill in the art. Polypeptides can also be used to raise antibodies, which in turn are used to measure protein expression from a recombinant cell, as a way of assessing transformation of the host cell. Moreover, the polypeptides of the present invention can be used to test the biological activities described herein.

Diagnostic Assays

The compounds of the present invention are useful for diagnosis, treatment, prevention and/or prognosis of various disorders in mammals, preferably humans. Such disorders include, but are not limited to, those described in the legends for Tables 1D and 1E and as indicated in the “Preferred Indications” columns in Table 1D and Table 1E; and, also as described herein under the section heading “Biological Activities”.

For a number of disorders, substantially altered (increased or decreased) levels of gene expression can be detected in tissues, cells or bodily fluids (e.g., sera, plasma, urine, semen, synovial fluid or spinal fluid) taken from an individual having such a disorder, relative to a “standard” gene expression level, that is, the expression level in tissues or bodily fluids from an individual not having the disorder. Thus, the invention provides a diagnostic method useful during diagnosis of a disorder, which involves measuring the expression level of the gene encoding the polypeptide in tissues, cells or body fluid from an individual and comparing the measured gene expression level with a standard gene expression level, whereby an increase or decrease in the gene expression level(s) compared to the standard is indicative of a disorder. These diagnostic assays may be performed in vivo or in vitro, such as, for example, on blood samples, biopsy tissue or autopsy tissue.

The present invention is also useful as a prognostic indicator, whereby patients exhibiting enhanced or depressed gene expression will experience a worse clinical outcome relative to patients expressing the gene at a level nearer the standard level.

In certain embodiments, a polypeptide of the invention, or polynucleotides, antibodies, agonists, or antagonists corresponding to that polypeptide, may be used to diagnose and/or prognose diseases and/or disorders associated with the tissue(s) in which the polypeptide of the invention is expressed, including one, two, three, four, five, or more tissues disclosed in Table 1B.2, column 5 (Tissue Distribution Library Code).

By “assaying the expression level of the gene encoding the polypeptide” is intended qualitatively or quantitatively measuring or estimating the level of the polypeptide of the invention or the level of the mRNA encoding the polypeptide of the invention in a first biological sample either directly (e.g., by determining or estimating absolute protein level or mRNA level) or relatively (e.g., by comparing to the polypeptide level or mRNA level in a second biological sample). Preferably, the polypeptide expression level or mRNA level in the first biological sample is measured or estimated and compared to a standard polypeptide level or mRNA level, the standard being taken from a second biological sample obtained from an individual not having the disorder or being determined by averaging levels from a population of individuals not having the disorder. As will be appreciated in the art, once a standard polypeptide level or mRNA level is known, it can be used repeatedly as a standard for comparison.

By “biological sample” is intended any biological sample obtained from an individual, cell line, tissue culture, or other source containing polypeptides of the invention (including portions thereof) or mRNA. As indicated, biological samples include body fluids (such as sera, plasma, urine, synovial fluid and spinal fluid) and tissue sources found to express the full length or fragments thereof of a polypeptide or mRNA. Methods for obtaining tissue biopsies and body fluids from mammals are well known in the art. Where the biological sample is to include mRNA, a tissue biopsy is the preferred source.

Total cellular RNA can be isolated from a biological sample using any suitable technique such as the single-step guanidinium-thiocyanate-phenol-chloroform method described in Chomczynski and Sacchi, Anal. Biochem. 162:156-159 (1987). Levels of mRNA encoding the polypeptides of the invention are then assayed using any appropriate method. These include Northern blot analysis, S1 nuclease mapping, the polymerase chain reaction (PCR), reverse transcription in combination with the polymerase chain reaction (RT-PCR), and reverse transcription in combination with the ligase chain reaction (RT-LCR).

The present invention also relates to diagnostic assays such as quantitative and diagnostic assays for detecting levels of polypeptides of the invention, in a biological sample (e.g., cells and tissues), including determination of normal and abnormal levels of polypeptides. Thus, for instance, a diagnostic assay in accordance with the invention for detecting over-expression of polypeptides of the invention compared to normal control tissue samples may be used to detect the presence of tumors. Assay techniques that can be used to determine levels of a polypeptide, such as a polypeptide of the present invention in a sample derived from a host are well-known to those of skill in the art. Such assay methods include radioimmunoassays, competitive-binding assays, Western Blot analysis and ELISA assays. Assaying polypeptide levels in a biological sample can occur using any art-known method.

Assaying polypeptide levels in a biological sample can occur using antibody-based techniques. For example, polypeptide expression in tissues can be studied with classical immunohistological methods (Jalkanen et al., J. Cell. Biol. 101:976-985 (1985); Jalkanen, M., et al., J. Cell. Biol. 105:3087-3096 (1987)). Other antibody-based methods useful for detecting polypeptide gene expression include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA). Suitable antibody assay labels are known in the art and include enzyme labels, such as, glucose oxidase, and radioisotopes, such as iodine (¹²⁵I, ¹²¹I), carbon (¹⁴C), sulfur (³⁵S), tritium (³H), indium (¹²¹In), and technetium (^(99m)Tc), and fluorescent labels, such as fluorescein and rhodamine, and biotin.

The tissue or cell type to be analyzed will generally include those which are known, or suspected, to express the gene of inteest (such as, for example, cancer). The protein isolation methods employed herein may, for example, be such as those described in Harlow and Lane (Harlow, E. and Lane, D., 1988, “Antibodies: A Laboratory Manual”, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.), which is incorporated herein by reference in its entirety. The isolated cells can be derived from cell culture or from a patient. The analysis of cells taken from culture may be a necessary step in the assessment of cells that could be used as part of a cell-based gene therapy technique or, alternatively, to test the effect of compounds on the expression of the gene.

For example, antibodies, or fragments of antibodies, such as those described herein, may be used to quantitatively or qualitatively detect the presence of gene products or conserved variants or peptide fragments thereof. This can be accomplished, for example, by immunofluorescence techniques employing a fluorescently labeled antibody coupled with light microscopic, flow cytometric, or fluorimetric detection.

In a preferred embodiment, antibodies, or fragments of antibodies directed to any one or all of the predicted epitope domains of the polypeptides of the invention (shown in column 7 of Table 1B.1) may be used to quantitatively or qualitatively detect the presence of gene products or conserved variants or peptide fragments thereof. This can be accomplished, for example, by immunofluorescence techniques employing a fluorescently labeled antibody coupled with light microscopic, flow cytometric, or fluorimetric detection.

In an additional preferred embodiment, antibodies, or fragments of antibodies directed to a conformational epitope of a polypeptide of the invention may be used to quantitatively or qualitatively detect the presence of gene products or conserved variants or peptide fragments thereof. This can be accomplished, for example, by immunofluorescence techniques employing a fluorescently labeled antibody coupled with light microscopic, flow cytometric, or fluorimetric detection.

The antibodies (or fragments thereof), and/or polypeptides of the present invention may, additionally, be employed histologically, as in immunofluorescence, immunoelectron microscopy or non-immunological assays, for in situ detection of gene products or conserved variants or peptide fragments thereof. In situ detection may be accomplished by removing a histological specimen from a patient, and applying thereto a labeled antibody or polypeptide of the present invention. The antibody (or fragment thereof) or polypeptide is preferably applied by overlaying the labeled antibody (or fragment) onto a biological sample. Through the use of such a procedure, it is possible to determine not only the presence of the gene product, or conserved variants or peptide fragments, or polypeptide binding, but also its distribution in the examined tissue. Using the present invention, those of ordinary skill will readily perceive that any of a wide variety of histological methods (such as staining procedures) can be modified in order to achieve such in situ detection.

Immunoassays and non-immunoassays for gene products or conserved variants or peptide fragments thereof will typically comprise incubating a sample, such as a biological fluid, a tissue extract, freshly harvested cells, or lysates of cells which have been incubated in cell culture, in the presence of a detectably labeled antibody capable of binding gene products or conserved variants or peptide fragments thereof, and detecting the bound antibody by any of a number of techniques well-known in the art.

The biological sample may be brought in contact with and immobilized onto a solid phase support or carrier such as nitrocellulose, or other solid support which is capable of immobilizing cells, cell particles or soluble proteins. The support may then be washed with suitable buffers followed by treatment with the detectably labeled antibody or detectable polypeptide of the invention. The solid phase support may then be washed with the buffer a second time to remove unbound antibody or polypeptide. Optionally the antibody is subsequently labeled. The amount of bound label on solid support may then be detected by conventional means.

By “solid phase support or carrier” is intended any support capable of binding an antigen or an antibody. Well-known supports or carriers include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylases, natural and modified celluloses, polyacrylamides, gabbros, and magnetite. The nature of the carrier can be either soluble to some extent or insoluble for the purposes of the present invention. The support material may have virtually any possible structural configuration so long as the coupled molecule is capable of binding to an antigen or antibody. Thus, the support configuration may be spherical, as in a bead, or cylindrical, as in the inside surface of a test tube, or the external surface of a rod. Alternatively, the surface may be flat such as a sheet, test strip, etc. Preferred supports include polystyrene beads. Those skilled in the art will know many other suitable carriers for binding antibody or antigen, or will be able to ascertain the same by use of routine experimentation.

The binding activity of a given lot of antibody or antigen polypeptide may be determined according to well known methods. Those skilled in the art will be able to determine operative and optimal assay conditions for each determination by employing routine experimentation.

In addition to assaying polypeptide levels or polynucleotide levels in a biological sample obtained from an individual, polypeptide or polynucleotide can also be detected in vivo by imaging. For example, in one embodiment of the invention, polypeptides and/or antibodies of the invention are used to image diseased cells, such as neoplasms. In another embodiment, polynucleotides of the invention (e.g., polynucleotides complementary to all or a portion of an mRNA) and/or antibodies (e.g., antibodies directed to any one or a combination of the epitopes of a polypeptide of the invention, antibodies directed to a conformational epitope of a polypeptide of the invention, or antibodies directed to the full length polypeptide expressed on the cell surface of a mammalian cell) are used to image diseased or neoplastic cells.

Antibody labels or markers for in vivo imaging of polypeptides of the invention include those detectable by X-radiography, NMR, MRI, CAT-scans or ESR. For X-radiography, suitable labels include radioisotopes such as barium or cesium, which emit detectable radiation but are not overtly harmful to the subject. Suitable markers for NMR and ESR include those with a detectable characteristic spin, such as deuterium, which may be incorporated into the antibody by labeling of nutrients for the relevant hybridoma. Where in vivo imaging is used to detect enhanced levels of polypeptides for diagnosis in humans, it may be preferable to use human antibodies or “humanized” chimeric monoclonal antibodies. Such antibodies can be produced using techniques described herein or otherwise known in the art. For example methods for producing chimeric antibodies are known in the art. See, for review, Morrison, Science 229:1202 (1985); Oi et al., BioTechniques 4:214 (1986); Cabilly et al., U.S. Pat. No. 4,816,567; Taniguchi et al., EP 171496; Morrison et al., EP 173494; Neuberger et al., WO 8601533; Robinson et al., WO 8702671; Boulianne et al., Nature 312:643 (1984); Neuberger et al., Nature 314:268 (1985).

Additionally, any polypeptides of the invention whose presence can be detected, can be administered. For example, polypeptides of the invention labeled with a radio-opaque or other appropriate compound can be administered and visualized in vivo, as discussed, above for labeled antibodies. Further, such polypeptides can be utilized for in vitro diagnostic procedures.

A polypeptide-specific antibody or antibody fragment which has been labeled with an appropriate detectable imaging moiety, such as a radioisotope (for example, ¹³¹I, ¹¹²In, ^(99m)Tc), a radio-opaque substance, or a material detectable by nuclear magnetic resonance, is introduced (for example, parenterally, subcutaneously or intraperitoneally) into the mammal to be examined for a disorder. It will be understood in the art that the size of the subject and the imaging system used will determine the quantity of imaging moiety needed to produce diagnostic images. In the case of a radioisotope moiety, for a human subject, the quantity of radioactivity injected will normally range from about 5 to 20 millicuries of ^(99m)Tc. The labeled antibody or antibody fragment will then preferentially accumulate at the location of cells which contain the antigenic protein. In vivo tumor imaging is described in S. W. Burchiel et al., “Immunopharmacokinetics of Radiolabeled Antibodies and Their Fragments” (Chapter 13 in Tumor Imaging: The Radiochemical Detection of Cancer, S. W. Burchiel and B. A. Rhodes, eds., Masson Publishing Inc. (1982)).

With respect to antibodies, one of the ways in which an antibody of the present invention can be detectably labeled is by linking the same to a reporter enzyme and using the linked product in an enzyme immunoassay (EIA) (Voller, A., “The Enzyme Linked Immunosorbent Assay (ELISA)”, 1978, Diagnostic Horizons 2:1-7, Microbiological Associates Quarterly Publication, Walkersville, Md.); Voller et al., J. Clin. Pathol. 31:507-520 (1978); Butler, J. E., Meth. Enzymol. 73:482-523 (1981); Maggio, E. (ed.), 1980, Enzyme Immunoassay, CRC Press, Boca Raton, Fla.,; Ishikawa, E. et al., (eds.), 1981, Enzyme Immunoassay, Kgaku Shoin, Tokyo). The reporter enzyme which is bound to the antibody will react with an appropriate substrate, preferably a chromogenic substrate, in such a manner as to produce a chemical moiety which can be detected, for example, by spectrophotometric, fluorimetric or by visual means. Reporter enzymes which can be used to detectably label the antibody include, but are not limited to, malate dehydrogenase, staphylococcal nuclease, delta-5-steroid isomerase, yeast alcohol dehydrogenase, alpha-glycerophosphate, dehydrogenase, triose phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase and acetylcholinesterase. Additionally, the detection can be accomplished by colorimetric methods which employ a chromogenic substrate for the reporter enzyme. Detection may also be accomplished by visual comparison of the extent of enzymatic reaction of a substrate in comparison with similarly prepared standards.

Detection may also be accomplished using any of a variety of other immunoassays. For example, by radioactively labeling the antibodies or antibody fragments, it is possible to detect polypeptides through the use of a radioimmunoassay (RIA) (see, for example, Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radioligand Assay Techniques, The Endocrine Society, March, 1986, which is incorporated by reference herein). The radioactive isotope can be detected by means including, but not limited to, a gamma counter, a scintillation counter, or autoradiography.

It is also possible to label the antibody with a fluorescent compound. When the fluorescently labeled antibody is exposed to light of the proper wave length, its presence can then be detected due to fluorescence. Among the most commonly used fluorescent labeling compounds are fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, allophycocyanin, ophthaldehyde and fluorescamine.

The antibody can also be detectably labeled using fluorescence emitting metals such as ¹⁵²Eu, or others of the lanthanide series. These metals can be attached to the antibody using such metal chelating groups as diethylenetriaminepentacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA).

The antibody also can be detectably labeled by coupling it to a chemiluminescent compound. The presence of the chemiluminescent-tagged antibody is then determined by detecting the presence of luminescence that arises during the course of a chemical reaction. Examples of particularly useful chemiluminescent labeling compounds are luminol, isoluminol, theromatic acridinium ester, imidazole, acridinium salt and oxalate ester.

Likewise, a bioluminescent compound may be used to label the antibody of the present invention. Bioluminescence is a type of chemiluminescence found in biological systems in, which a catalytic protein increases the efficiency of the chemiluminescent reaction. The presence of a bioluminescent protein is determined by detecting the presence of luminescence. Important bioluminescent compounds for purposes of labeling are luciferin, luciferase and aequorin.

Methods for Detecting Diseases

In general, a disease may be detected in a patient based on the presence of one or more proteins of the invention and/or polynucleotides encoding such proteins in a biological sample (for example, blood, sera, urine, and/or tumor biopsies) obtained from the patient. In other words, such proteins may be used as markers to indicate the presence or absence of a disease or disorder, including cancer and/or as described elsewhere herein. In addition, such proteins may be useful for the detection of other diseases and cancers. The binding agents provided herein generally permit detection of the level of antigen that binds to the agent in the biological sample. Polynucleotide primers and probes may be used to detect the level of mRNA encoding polypeptides of the invention, which is also indicative of the presence or absence of a disease or disorder, including cancer. In general, polypeptides of the invention should be present at a level that is at least three fold higher in diseased tissue than in normal tissue.

There are a variety of assay formats known to those of ordinary skill in the art for using a binding agent to detect polypeptide markers in a sample. See, e.g., Harlow and Lane, supra. In general, the presence or absence of a disease in a patient may be determined by (a) contacting a biological sample obtained from a patient with a binding agent; (b) detecting in the sample a level of polypeptide that binds to the binding agent; and (c) comparing the level of polypeptide with a predetermined cut-off value.

In a preferred embodiment, the assay involves the use of a binding agent(s) immobilized on a solid support to bind to and remove the polypeptide of the invention from the remainder of the sample. The bound polypeptide may then be detected using a detection reagent that contains a reporter group and specifically binds to the binding agent/polypeptide complex. Such detection reagents may comprise, for example, a binding agent that specifically binds to the polypeptide or an antibody or other agent that specifically binds to the binding agent, such as an anti-immunoglobulin, protein G, protein A or a lectin. Alternatively, a competitive assay may be utilized, in which a polypeptide is labeled with a reporter group and allowed to bind to the immobilized binding agent after incubation of the binding agent with the sample. The extent to which components of the sample inhibit the binding of the labeled polypeptide to the binding agent is indicative of the reactivity of the sample with the immobilized binding agent. Suitable polypeptides for use within such assays include polypeptides of the invention and portions thereof, or antibodies, to which the binding agent binds, as described above.

The solid support may be any material known to those of skill in the art to which polypeptides of the invention may be attached. For example, the solid support may be a test well in a microtiter plate or a nitrocellulose or other suitable membrane. Alternatively, the support may be a bead or disc, such as glass fiberglass, latex or a plastic material such as polystyrene or polyvinylchloride. The support may also be a magnetic particle or a fiber optic sensor, such as those disclosed, for example, in U.S. Pat. No. 5,359,681. The binding agent may be immobilized on the solid support using a variety of techniques known to those of skill in the art, which are amply described in the patent and scientific literature. In the context of the present invention, the term “immobilization” refers to both noncovalent association, such as adsorption, and covalent attachment (which may be a direct linkage between the agent and functional groups on the support or may be a linkage by way of a cross-linking agent). Immobilization by adsorption to a well in a microtiter plate or to a membrane is preferred. In such cases, adsorption may be achieved by contacting the binding agent, in a suitable buffer, with the solid support for the suitable amount of time. The contact time varies with temperature, but is typically between about 1 hour and about 1 day. In general, contacting a well of plastic microtiter plate (such as polystyrene or polyvinylchloride) with an amount of binding agent ranging from about 10 ng to about 10 ug, and preferably about 100 ng to about 1 ug, is sufficient to immobilize an adequate amount of binding agent.

Covalent attachment of binding agent to a solid support may generally be achieved by first reacting the support with a bifunctional reagent that will react with both the support and a functional group, such as a hydroxyl or amino group, on the binding agent. For example, the binding agent may be covalently attached to supports having an appropriate polymer coating using benzoquinone or by condensation of an aldehyde group on the support with an amine and an active hydrogen on the binding partner (see, e.g., Pierce Immunotechnology Catalog and Handbook, 1991, at A12-A13).

Gene Therapy Methods

Also encompassed by the invention are gene therapy methods for treating or preventing disorders, diseases and conditions. The gene therapy methods relate to the introduction of nucleic acid (DNA, RNA and antisense DNA or RNA) sequences into an animal to achieve expression of the polypeptide of the present invention. This method requires a polynucleotide which codes for a polypeptide of the present invention operatively linked to a promoter and any other genetic elements necessary for the expression of the polypeptide by the target tissue. Such gene therapy and delivery techniques are known in the art, see, for example, WO90/11092, which is herein incorporated by reference.

Thus, for example, cells from a patient may be engineered with a polynucleotide (DNA or RNA) comprising a promoter operably linked to a polynucleotide of the present invention ex vivo, with the engineered cells then being provided to a patient to be treated with the polypeptide of the present invention. Such methods are well-known in the art. For example, see Belldegrun, A., et al., J. Natl. Cancer Inst. 85: 207-216 (1993); FerTantini, M. et al., Cancer Research 53: 1107-1112 (1993); FelTantini, M. et al., J. Immunology 153: 4604-4615 (1994); Kaido, T., et al., Int. J. Cancer 60: 221-229 (1995); Ogura, H., et al., Cancer Research 50: 5102-5106 (1990); Santodonato, L., et al., Human Gene Therapy 7:1-10 (1996); Santodonato, L., et al., Gene Therapy 4:1246-1255 (1997); and Zhang, J.-F. et al., Cancer Gene Therapy 3: 31-38 (1996)), which are herein incorporated by reference. In one embodiment, the cells which are engineered are arterial cells. The arterial cells may be reintroduced into the patient through direct injection to the artery, the tissues surrounding the artery, or through catheter injection.

As discussed in more detail below, the polynucleotide constructs can be delivered by any method that delivers injectable materials to the cells of an animal, such as, injection into the interstitial space of tissues (heart, muscle, skin, lung, liver, and the like). The polynucleotide constructs may be delivered in a pharmaceutically acceptable liquid or aqueous carrier.

In one embodiment, the polynucleotide of the present invention is delivered as a naked polynucleotide. The term “naked” polynucleotide, DNA or RNA refers to sequences that are free from any delivery vehicle that acts to assist, promote or facilitate entry into the cell, including viral sequences, viral particles, liposome formulations, lipofectin or precipitating agents and the like. However, the polynucleotide of the present invention can also be delivered in liposome formulations and lipofectin formulations and the like can be prepared by methods well known to those skilled in the art. Such methods are described, for example, in U.S. Pat. Nos. 5,593,972, 5,589,466, and 5,580,859, which are herein incorporated by reference.

The polynucleotide vector constructs used in the gene therapy method are preferably constructs that will not integrate into the host genome nor will they contain sequences that allow for replication. Appropriate vectors include pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; pSVK3, pBPV, pMSG and pSVL available from Pharmacia; and pEF1/V5, pcDNA3.1, and pRc/CMV2 available from Invitrogen. Other suitable vectors will be readily apparent to the skilled artisan.

Any strong promoter known to those skilled in the art can be used for driving the expression of the polynucleotide sequence. Suitable promoters include adenoviral promoters, such as the adenoviral major late promoter; or heterologous promoters, such as the cytomegalovirus (CMV) promoter; the respiratory syncytial virus (RSV) promoter; inducible promoters, such as the MMT promoter, the metallothionein promoter; heat shock promoters; the albumin promoter; the ApoAI promoter; human globin promoters; viral thymidine kinase promoters, such as the Herpes Simplex thymidine kinase promoter; retroviral LTRs; the b-actin promoter; and human growth hormone promoters. The promoter also may be the native promoter for the polynucleotide of the present invention.

Unlike other gene therapy techniques, one major advantage of introducing naked nucleic acid sequences into target cells is the transitory nature of the polynucleotide synthesis in the cells. Studies have shown that non-replicating DNA sequences can be introduced into cells to provide production of the desired polypeptide for periods of up to six months.

The polynucleotide construct can be delivered to the interstitial space of tissues within the an animal, including of muscle, skin, brain, lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone, cartilage, pancreas, kidney, gall bladder, stomach, intestine, testis, ovary, uterus, rectum, nervous system, eye, gland, and connective tissue. Interstitial space of the tissues comprises the intercellular, fluid, mucopolysaccharide matrix among the reticular fibers of organ tissues, elastic fibers in the walls of vessels or chambers, collagen fibers of fibrous tissues, or that same matrix within connective tissue ensheathing muscle cells or in the lacunae of bone. It is similarly the space occupied by the plasma of the circulation and the lymph fluid of the lymphatic channels. Delivery to the interstitial space of muscle tissue is preferred for the reasons discussed below. They may be conveniently delivered by injection into the tissues comprising these cells. They are preferably delivered to and expressed in persistent, non-dividing cells which are differentiated, although delivery and expression may be achieved in non-differentiated or less completely differentiated cells, such as, for example, stem cells of blood or skin fibroblasts. In vivo muscle cells are particularly competent in their ability to take up and express polynucleotides.

For the naked nucleic acid sequence injection, an effective dosage amount of DNA or RNA will be in the range of from about 0.05 mg/kg body weight to about 50 mg/kg body weight. Preferably the dosage will be from about 0.005 mg/kg to about 20 mg/kg and more preferably from about 0.05 mg/kg to about 5 mg/kg. Of course, as the artisan of ordinary skill will appreciate, this dosage will vary according to the tissue site of injection. The appropriate and effective dosage of nucleic acid sequence can readily be determined by those of ordinary skill in the art and may depend on the condition being treated and the route of administration.

The preferred route of administration is by the parenteral route of injection into the interstitial space of tissues. However, other parenteral routes may also be used, such as, inhalation of an aerosol formulation particularly for delivery to lungs or bronchial tissues, throat or mucous membranes of the nose. In addition, naked DNA constructs can be delivered to arteries during angioplasty by the catheter used in the procedure.

The naked polynucleotides are delivered by any method known in the art, including, but not limited to, direct needle injection at the delivery site, intravenous injection, topical administration, catheter infusion, and so-called “gene guns”. These delivery methods are known in the art.

The constructs may also be delivered with delivery vehicles such as viral sequences, viral particles, liposome formulations, lipofectin, precipitating agents, etc. Such methods of delivery are known in the art.

In certain embodiments, the polynucleotide constructs are complexed in a liposome preparation. Liposomal preparations for use in the instant invention include cationic (positively charged), anionic (negatively charged) and neutral preparations. However, cationic liposomes are particularly preferred because a tight charge complex can be formed between the cationic liposome and the polyanionic nucleic acid. Cationic liposomes have been shown to mediate intracellular delivery of plasmid DNA (Feigner et al., Proc. Natl. Acad. Sci. USA (1987) 84:7413-7416, which is herein incorporated by reference); mRNA (Malone et al., Proc. Natl. Acad. Sci. USA (1989) 86:6077-6081, which is herein incorporated by reference); and purified transcription factors (Debs et al., J. Biol. Chem. (1990) 265:10189-10192, which is herein incorporated by reference), in functional form.

Cationic liposomes are readily available. For example, N[1-2,3-dioleyloxy)propyl]-N,N,N-triethylammonium (DOTMA) liposomes are particularly useful and are available under the trademark Lipofectin, from GIBCO BRL, Grand Island, N.Y. (See, also, Feigner et al., Proc. Natl. Acad. Sci. USA (1987) 84:7413-7416, which is herein incorporated by reference). Other commercially available liposomes include transfectace (DDAB/DOPE) and DOTAP/DOPE (Boehringer).

Other cationic liposomes can be prepared from readily available materials using techniques well known in the art. See, e.g. PCT Publication No. WO 90/11092 (which is herein incorporated by reference) for a description of the synthesis of DOTAP (1,2-bis(oleoyloxy)-3-(trimethylammonio)propane) liposomes. Preparation of DOTMA liposomes is explained in the literature, see, e.g., P. Feigner et al., Proc. Natl. Acad. Sci. USA 84:7413-7417, which is herein incorporated by reference. Similar methods can be used to prepare liposomes from other cationic lipid materials.

Similarly, anionic and neutral liposomes are readily available, such as from Avanti Polar Lipids (Birmingham, Ala.), or can be easily prepared using readily available materials. Such materials include phosphatidyl, choline, cholesterol, phosphatidyl ethanolamine, dioleoylphosphatidyl choline (DOPC), dioleoylphosphatidyl glycerol (DOPG), dioleoylphoshatidyl ethanolamine (DOPE), among others. These materials can also be mixed with the DOTMA and DOTAP starting materials in appropriate ratios. Methods for making liposomes using these materials are well known in the art.

For example, commercially dioleoylphosphatidyl choline (DOPC), dioleoylphosphatidyl glycerol (DOPG), and dioleoylphosphatidyl ethanolamine (DOPE) can be used in various combinations to make conventional liposomes, with or without the addition of cholesterol. Thus, for example, DOPG/DOPC vesicles can be prepared by drying 50 mg each of DOPG and DOPC under a stream of nitrogen gas into a sonication vial. The sample is placed under a vacuum pump overnight and is hydrated the following day with deionized water. The sample is then sonicated for 2 hours in a capped vial, using a Heat Systems model 350 sonicator equipped with an inverted cup (bath type) probe at the maximum setting while the bath is circulated at 15EC. Alternatively, negatively charged vesicles can be prepared without sonication to produce multilamellar vesicles or by extrusion through nucleopore membranes to produce unilamellar vesicles of discrete size. Other methods are known and available to those of skill in the art.

The liposomes can comprise multilamellar vesicles (MLVs), small unilamellar vesicles (SUVs), or large unilamellar vesicles (LUVs), with SUVs being preferred. The various liposome-nucleic acid complexes are prepared using methods well known in the art. See, e.g., Straubinger et al., Methods of Immunology (1983), 101:512-527, which is herein incorporated by reference. For example, MLVs containing nucleic acid can be prepared by depositing a thin film of phospholipid on the walls of a glass tube and subsequently hydrating with a solution of the material to be encapsulated. SUVs are prepared by extended sonication of MLVs to produce a homogeneous population of unilamellar liposomes. The material to be entrapped is added to a suspension of preformed MLVs and then sonicated. When using liposomes containing cationic lipids, the dried lipid film is resuspended in an appropriate solution such as sterile water or an isotonic buffer solution such as 10 mM Tris/NaCl, sonicated, and then the preformed liposomes are mixed directly with the DNA. The liposome and DNA form a very stable complex due to binding of the positively charged liposomes to the cationic DNA. SUVs find use with small nucleic acid fragments. LUVs are prepared by a number of methods, well known in the art. Commonly used methods include Ca²⁺-EDTA chelation (Papahadjopoulos et al., Biochim. Biophys. Acta (1975) 394:483; Wilson et al., Cell 17:77 (1979)); ether injection (Deamer, D. and Bangham, A., Biochim. Biophys. Acta 443:629 (1976); Ostro et al., Biochem. Biophys. Res. Commun. 76:836 (1977); Fraley et al., Proc. Natl. Acad. Sci. USA 76:3348 (1979)); detergent dialysis (Enoch, H. and Strittmatter, P., Proc. Natl. Acad. Sci. USA 76:145 (1979)); and reverse-phase evaporation (REV) (Fraley et al., J. Biol. Chem. 255:10431 (1980); Szoka, F. and Papahadjopoulos, D., Proc. Natl. Acad. Sci. USA 75:145 (1978); Schaefer-Ridder et al., Science 215:166 (1982)), which are herein incorporated by reference.

Generally, the ratio of DNA to liposomes will be from about 10:1 to about 1:10. Preferably, the ration will be from about 5:1 to about 1:5. More preferably, the ration will be about 3:1 to about 1:3. Still more preferably, the ratio will be about 1:1.

U.S. Pat. No. 5,676,954 (which is herein incorporated by reference) reports on the injection of genetic material, complexed with cationic liposomes carriers, into mice. U.S. Pat. Nos. 4,897,355, 4,946,787, 5,049,386, 5,459,127, 5,589,466, 5,693,622, 5,580,859, 5,703,055, and international publication no. WO 94/9469 (which are herein incorporated by reference) provide cationic lipids for use in transfecting DNA into cells and mammals. U.S. Pat. Nos. 5,589,466, 5,693,622, 5,580,859, 5,703,055, and international publication no. WO 94/9469 provide methods for delivering DNA-cationic lipid complexes to mammals.

In certain embodiments, cells are engineered, ex vivo or in vivo, using a retroviral particle containing RNA which comprises a sequence encoding a polypeptide of the present invention. Retroviruses from which the retroviral plasmid vectors may be derived include, but are not limited to, Moloney Murine Leukemia Virus, spleen necrosis virus, Rous sarcoma Virus, Harvey Sarcoma Virus, avian leukosis virus, gibbon ape leukemia virus, human immunodeficiency virus, Myeloproliferative Sarcoma Virus, and mammary tumor virus.

The retroviral plasmid vector is employed to transduce packaging cell lines to form producer cell lines. Examples of packaging cells which may be transfected include, but are not limited to, the PE501, PA317, R-2, R-AM, PA12, T19-14X, VT-19-17-H2, RCRE, RCRIP, GP+E-86, GP+envAm12, and DAN cell lines as described in Miller, Human Gene Therapy 1:5-14 (1990), which is incorporated herein by reference in its entirety. The vector may transduce the packaging cells through any means known in the art. Such means include, but are not limited to, electroporation, the use of liposomes, and CaPO₄ precipitation. In one alternative, the retroviral plasmid vector may be encapsulated into a liposome, or coupled to a lipid, and then administered to a host.

The producer cell line generates infectious retroviral vector particles which include polynucleotide encoding a polypeptide of the present invention. Such retroviral vector particles then may be employed, to transduce eukaryotic cells, either in vitro or in vivo. The transduced eukaryotic cells will express a polypeptide of the present invention.

In certain other embodiments, cells are engineered, ex vivo or in vivo, with polynucleotide contained in an adenovirus vector. Adenovirus can be manipulated such that it encodes and expresses a polypeptide of the present invention, and at the same time is inactivated in terms of its ability to replicate in a normal lytic viral life cycle. Adenovirus expression is achieved without integration of the viral DNA into the host cell chromosome, thereby alleviating concerns about insertional mutagenesis. Furthermore, adenoviruses have been used as live enteric vaccines for many years with an excellent safety profile (Schwartz et al. Am. Rev. Respir. Dis. 109:233-238 (1974)). Finally, adenovirus mediated gene transfer has been demonstrated in a number of instances including transfer of alpha-1-antitrypsin and CFTR to the lungs of cotton rats (Rosenfeld, M. A. et al. (1991) Science 252:431-434; Rosenfeld et al., (1992) Cell 68:143-155). Furthermore, extensive studies to attempt to establish adenovirus as a causative agent in human cancer were uniformly negative (Green, M. et al. (1979) Proc. Natl. Acad. Sci. USA 76:6606).

Suitable adenoviral vectors useful in the present invention are described, for example, in Kozarsky and Wilson, Curr. Opin. Genet. Devel. 3:499-503 (1993); Rosenfeld et al., Cell 68:143-155 (1992); Engelhardt et al., Human Genet. Ther. 4:759-769 (1993); Yang et al., Nature Genet. 7:362-369 (1994); Wilson et al., Nature 365:691-692 (1993); and U.S. Pat. No. 5,652,224, which are herein incorporated by reference. For example, the adenovirus vector Ad2 is useful and can be grown in human 293 cells. These cells contain the E1 region of adenovirus and constitutively express E1a and E1b, which complement the defective adenoviruses by providing the products of the genes deleted from the vector. In addition to Ad2, other varieties of adenovirus (e.g., Ad3, Ad5, and Ad7) are also useful in the present invention.

Preferably, the adenoviruses used in the present invention are replication deficient. Replication deficient adenoviruses require the aid of a helper virus and/or packaging cell line to form infectious particles. The resulting virus is capable of infecting cells and can express a polynucleotide of interest which is operably linked to a promoter, but cannot replicate in most cells. Replication deficient adenoviruses may be deleted in one or more of all or a portion of the following genes: E1a, E1b, E3, E4, E2a, or L1 through L5.

In certain other embodiments, the cells are engineered, cx vivo or in vivo, using an adeno-associated virus (AAV). AAVs are naturally occurring defective viruses that require helper viruses to produce infectious particles (Muzyczka, N., Curr. Topics in Microbiol. Immunol. 158:97 (1992)). It is also one of the few viruses that may integrate its DNA into non-dividing cells. Vectors containing as little as 300 base pairs of AAV can be packaged and can integrate, but space for exogenous DNA is limited to about 4.5 kb. Methods for producing and using such AAVs are known in the art. See, for example, U.S. Pat. Nos. 5,139,941, 5,173,414, 5,354,678, 5,436,146, 5,474,935, 5,478,745, and 5,589,377.

For example, an appropriate AAV vector for use in the present invention will include all the sequences necessary for DNA replication, encapsidation, and host-cell integration. The polynucleotide construct is inserted into the AAV vector using standard cloning methods, such as those found in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989). The recombinant AAV vector is then transfected into packaging cells which are infected with a helper virus, using any standard technique, including lipofection, electroporation, calcium phosphate precipitation, etc. Appropriate helper viruses include adenoviruses, cytomegaloviruses, vaccinia viruses, or herpes viruses. Once the packaging cells are transfected and infected, they will produce infectious AAV viral particles which contain the polynucleotide construct. These viral particles are then used to transduce eukaryotic cells, either ex vivo or in vivo. The transduced cells will contain the polynucleotide construct integrated into its genome, and will express a polypeptide of the invention.

Another method of gene therapy involves operably associating heterologous control regions and endogenous polynucleotide sequences (e.g. encoding a polypeptide of the present invention) via homologous recombination (see, e.g., U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication No. WO 96/29411, published Sep. 26, 1996; International Publication No. WO 94/12650, published Aug. 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), which are herein incorporated by reference. This method involves the activation of a gene which is present in the target cells, but which is not normally expressed in the cells, or is expressed at a lower level than desired.

Polynucleotide constructs are made, using standard techniques known in the art, which contain the promoter with targeting sequences flanking the promoter. Suitable promoters are described herein. The targeting sequence is sufficiently complementary to an endogenous sequence to permit homologous recombination of the promoter-targeting sequence with the endogenous sequence. The targeting sequence will be sufficiently near the 5′ end of the desired endogenous polynucleotide sequence so the promoter will be operably linked to the endogenous sequence upon homologous recombination.

The promoter and the targeting sequences can be amplified using PCR. Preferably, the amplified promoter contains distinct restriction enzyme sites on the 5′ and 3′ ends. Preferably, the 3′ end of the first targeting sequence contains the same restriction enzyme site as the 5′ end of the amplified promoter and the 5′ end of the second targeting sequence contains the same restriction site as the 3′ end of the amplified promoter. The amplified promoter and targeting sequences are digested and ligated together.

The promoter-targeting sequence construct is delivered to the cells, either as naked polynucleotide, or in conjunction with transfection-facilitating agents, such as liposomes, viral sequences, viral particles, whole viruses, lipofection, precipitating agents, etc., described in more detail above. The P promoter-targeting sequence can be delivered by any method, included direct needle injection, intravenous injection, topical administration, catheter infusion, particle accelerators, etc. The methods are described in more detail below.

The promoter-targeting sequence construct is taken up by cells. Homologous recombination between the construct and the endogenous sequence takes place, such that an endogenous sequence is placed under the control of the promoter. The promoter then drives the expression of the endogenous sequence.

The polynucleotide encoding a polypeptide of the present invention may contain a secretory signal sequence that facilitates secretion of the protein. Typically, the signal sequence is positioned in the coding region of the polynucleotide to be expressed towards or at the 5′ end of the coding region. The signal sequence may be homologous or heterologous to the polynucleotide of interest and may be homologous or heterologous to the cells to be transfected. Additionally, the signal sequence may be chemically synthesized using methods known in the art.

Any mode of administration of any of the above-described polynucleotides constructs can be used so long as the mode results in the expression of one or more molecules in an amount sufficient to provide a therapeutic effect. This includes direct needle injection, systemic injection, catheter infusion, biolistic injectors, particle accelerators (i.e., “gene guns”), gelfoam sponge depots, other commercially available depot materials, osmotic pumps (e.g., Alza minipumps), oral or suppositorial solid (tablet or pill) pharmaceutical formulations, and decanting or topical applications during surgery. For example, direct injection of naked calcium phosphate-precipitated plasmid into rat liver and rat spleen or a protein-coated plasmid into the portal vein has resulted in gene expression of the foreign gene in the rat livers (Kaneda et al., Science 243:375 (1989)).

A preferred method of local administration is by direct injection. Preferably, a recombinant molecule of the present invention complexed with a delivery vehicle is administered by direct injection into or locally within the area of arteries. Administration of a composition locally within the area of arteries refers to injecting the composition centimeters and preferably, millimeters within arteries.

Another method of local administration is to contact a polynucleotide construct of the present invention in or around a surgical wound. For example, a patient can undergo surgery and the polynucleotide construct can be coated on the surface of tissue inside the wound or the construct can be injected into areas of tissue inside the wound.

Therapeutic compositions useful in systemic administration, include recombinant molecules of the present invention complexed to a targeted delivery vehicle of the present invention. Suitable delivery vehicles for use with systemic administration comprise liposomes comprising ligands for targeting the vehicle to a particular site. In specific embodiments, suitable delivery vehicles for use with systemic administration comprise liposomes comprising polypeptides of the invention for targeting the vehicle to a particular site.

Preferred methods of systemic administration, include intravenous injection, aerosol, oral and percutaneous (topical) delivery. Intravenous injections can be performed using methods standard in the art. Aerosol delivery can also be performed using methods standard in the art (see, for example, Stribling et al., Proc. Natl. Acad. Sci. USA 189:11277-11281, 1992, which is incorporated herein by reference). Oral delivery can be performed by complexing a polynucleotide construct of the present invention to a carrier capable of withstanding degradation by digestive enzymes in the gut of an animal. Examples of such carriers, include plastic capsules or tablets, such as those known in the art. Topical delivery can be performed by mixing a polynucleotide construct of the present invention with a lipophilic reagent (e.g., DMSO) that is capable of passing into the skin.

Determining an effective amount of substance to be delivered can depend upon a number of factors including, for example, the chemical structure and biological activity of the substance, the age and weight of the animal, the precise condition requiring treatment and its severity, and the route of administration. The frequency of treatments depends upon a number of factors, such as the amount of polynucleotide constructs administered per dose, as well as the health and history of the subject. The precise amount, number of doses, and timing of doses will be determined by the attending physician or veterinarian.

Therapeutic compositions of the present invention can be administered to any animal, preferably to mammals and birds. Preferred mammals include humans, dogs, cats, mice, rats, rabbits sheep, cattle, horses and pigs, with humans being particularly preferred.

Biological Activities

Polynucleotides or polypeptides, or agonists or antagonists of the present invention, can be used in assays to test for one or more biological activities. If these polynucleotides or polypeptides, or agonists or antagonists of the present invention, do exhibit activity in a particular assay, it is likely that these molecules may be involved in the diseases associated with the biological activity. Thus, the polynucleotides and polypeptides, and agonists or antagonists could be used to treat the associated disease.

Members of the secreted family of proteins are believed to be involved in biological activities associated with, for example, cellular signaling. Accordingly, compositions of the invention (including polynucleotides, polypeptides and antibodies of the invention, and fragments and variants thereof) may be used in diagnosis, prognosis, prevention and/or treatment of diseases and/or disorders associated with aberrant activity of secreted polypeptides.

In preferred embodiments, compositions of the invention (including polynucleotides, polypeptides and antibodies of the invention, and fragments and variants thereof) may be used in the diagnosis, prognosis, prevention and/or treatment of diseases and/or disorders relating to diseases and disorders of the endocrine system, the nervous system (See, for example, “Neurological Disorders” section below), and the immune system (See, for example, “Immune Activity” section below).

In certain embodiments, a polypeptide of the invention, or polynucleotides, antibodies, agonists, or antagonists corresponding to that polypeptide, may be used to diagnose and/or prognose diseases and/or disorders associated with the tissue(s) in which the polypeptide of the invention is expressed including one, two, three, four, five, or more tissues disclosed in Table 1B.2, column 5 (Tissue Distribution Library Code).

Thus, polynucleotides, translation products and antibodies of the invention are useful in the diagnosis, detection and/or treatment of diseases and/or disorders associated with activities that include, but are not limited to, prohormone activation, neurotransmitter activity, cellular signaling, cellular proliferation, cellular differentiation, and cell migration.

More generally, polynucleotides, translation products and antibodies corresponding to this gene may be useful for the diagnosis, prognosis, prevention and/or treatment of diseases and/or disorders associated with the following systems.

Immune Activity

Polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, diagnosing and/or prognosing diseases, disorders, and/or conditions of the immune system, by, for example, activating or inhibiting the proliferation, differentiation, or mobilization (chemotaxis) of immune cells. Immune cells develop through a process called hematopoiesis, producing myeloid (platelets, red blood cells, neutrophils, and macrophages) and lymphoid (B and T lymphocytes) cells from pluripotent stem cells. The etiology of these immune diseases, disorders, and/or conditions may be genetic, somatic, such as cancer and some autoimmune diseases, acquired (e.g., by chemotherapy or toxins), or infectious. Moreover, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention can be used as a marker or detector of a particular immune system disease or disorder.

In another embodiment, a polypeptide of the invention, or polynucleotides, antibodies, agonists, or antagonists corresponding to that polypeptide, may be used to treat diseases and disorders of the immune system and/or to inhibit or enhance an immune response generated by cells associated with the tissue(s) in which the polypeptide of the invention is expressed, including one, two, three, four, five, or more tissues disclosed in Table 1B.2, column 5 (Tissue Distribution Library Code).

Polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, diagnosing, and/or prognosing immunodeficiencies, including both congenital and acquired immunodeficiencies. Examples of B cell immunodeficiencies in which immunoglobulin levels B cell function and/or B cell numbers are decreased include: X-linked agammaglobulinemia (Bruton's disease), X-linked infantile agammaglobulinemia, X-linked immunodeficiency with hyper IgM, non X-linked immunodeficiency with hyper IgM, X-linked lymphoproliferative syndrome (XLP), agammaglobulinemia including congenital and acquired agammaglobulinemia, adult onset agammaglobulinemia, late-onset agammaglobulinemia, dysgammaglobulinemia, hypogammaglobulinemia, unspecified hypogammaglobulinemia, recessive agammaglobulinemia (Swiss type), Selective IgM deficiency, selective IgA deficiency, selective IgG subclass deficiencies, IgG subclass deficiency (with or without IgA deficiency), Ig deficiency with increased IgM, IgG and IgA deficiency with increased IgM, antibody deficiency with normal or elevated Igs, Ig heavy chain deletions, kappa chain deficiency, B cell lymphoproliferative disorder (BLPD), common variable immunodeficiency (CVID), common variable immunodeficiency (CVI) (acquired), and transient hypogammaglobulinemia of infancy.

In specific embodiments, ataxia-telangiectasia or conditions associated with ataxia-telangiectasia are treated, prevented, diagnosed, and/or prognosing using the polypeptides or polynucleotides of the invention, and/or agonists or antagonists thereof.

Examples of congenital immunodeficiencies in which T cell and/or B cell function and/or number is decreased include, but are not limited to: DiGeorge anomaly, severe combined immunodeficiencies (SCID) (including, but not limited to, X-linked SCID, autosomal recessive SCID, adenosine deaminase deficiency, purine nucleoside phosphorylase (PNP) deficiency, Class II MHC deficiency (Bare lymphocyte syndrome), Wiskott-Aldrich syndrome, and ataxia telangiectasia), thymic hypoplasia, third and fourth pharyngeal pouch syndrome, 22q11.2 deletion, chronic mucocutaneous candidiasis, natural killer cell deficiency (NK), idiopathic CD4+ T-lymphocytopenia, immunodeficiency with predominant T cell defect (unspecified), and unspecified immunodeficiency of cell mediated immunity.

In specific embodiments, DiGeorge anomaly or conditions associated with DiGeorge anomaly are treated, prevented, diagnosed, and/or prognosed using polypeptides or polynucleotides of the invention, or antagonists or agonists thereof.

Other immunodeficiencies that may be treated, prevented, diagnosed, and/or prognosed using polypeptides or polynucleotides of the invention, and/or agonists or antagonists thereof, include, but are not limited to, chronic granulomatous disease, Chédiak-Higashi syndrome, myeloperoxidase deficiency, leukocyte glucose-6-phosphate dehydrogenase deficiency, X-linked lymphoproliferative syndrome (XLP), leukocyte adhesion deficiency, complement component deficiencies (including C1, C2, C3, C4, C5, C6, C7, C8 and/or C9 deficiencies), reticular dysgenesis, thymic alymphoplasia-aplasia, immunodeficiency with thymoma, severe congenital leukopenia, dysplasia with immunodeficiency, neonatal neutropenia, short limbed dwarfism, and Nezelof syndrome-combined immunodeficiency with Igs.

In a preferred embodiment, the immunodeficiencies and/or conditions associated with the immunodeficiencies recited above are treated, prevented, diagnosed and/or prognosed using polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention.

In a preferred embodiment polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention could be used as an agent to boost immunoresponsiveness among immunodeficient individuals. In specific embodiments, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention could be used as an agent to boost immunoresponsiveness among B cell and/or T cell immunodeficient individuals.

The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, diagnosing and/or prognosing autoimmune disorders. Many autoimmune disorders result from inappropriate recognition of self as foreign material by immune cells. This inappropriate recognition results in an immune response leading to the destruction of the host tissue. Therefore, the administration of polynucleotides and polypeptides of the invention that can inhibit an immune response, particularly the proliferation, differentiation, or chemotaxis of T-cells, may be an effective therapy in preventing autoimmune disorders.

Autoimmune diseases or disorders that may be treated, prevented, diagnosed and/or prognosed by polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention include, but are not limited to, one or more of the following: systemic lupus erythematosus, rheumatoid arthritis, ankylosing spondylitis, multiple sclerosis, autoimmune thyroiditis, Hashimoto's thyroiditis, autoimmune hemolytic anemia, hemolytic anemia, thrombocytopenia, autoimmune thrombocytopenia purpura, autoimmune neonatal thrombocytopenia, idiopathic thrombocytopenia purpura, purpura (e.g., Henloch-Scoenlein purpura), autoimmunocytopenia, Goodpasture's syndrome, Pemphigus vulgaris, myasthenia gravis, Grave's disease (hyperthyroidism), and insulin-resistant diabetes mellitus.

Additional disorders that are likely to have an autoimmune component that may be treated, prevented, and/or diagnosed with the compositions of the invention include, but are not limited to, type II collagen-induced arthritis, antiphospholipid syndrome, dermatitis, allergic encephalomyelitis, myocarditis, relapsing polychondritis, rheumatic heart disease, neuritis, uveitis ophthalmia, polyendocrinopathies, Reiter's Disease, Stiff-Man Syndrome, autoimmune pulmonary inflammation, autism, Guillain-Barre Syndrome, insulin dependent diabetes mellitus, and autoimmune inflammatory eye disorders.

Additional disorders that are likely to have an autoimmune component that may be treated, prevented, diagnosed and/or prognosed with the compositions of the invention include, but are not limited to, scleroderma with anti-collagen antibodies (often characterized, e.g., by nucleolar and other nuclear antibodies), mixed connective tissue disease (often characterized, e.g., by antibodies to extractable nuclear antigens (e.g., ribonucleoprotein)), polymyositis (often characterized, e.g., by nonhistone ANA), pernicious anemia (often characterized, e.g., by antiparietal cell, microsomes, and intrinsic factor antibodies), idiopathic Addison's disease (often characterized, e.g., by humoral and cell-mediated adrenal cytotoxicity, infertility (often characterized, e.g., by antispermatozoal antibodies), glomerulonephritis (often characterized, e.g., by glomerular basement membrane antibodies or immune complexes), bullous pemphigoid (often characterized, e.g., by IgG and complement in basement membrane), Sjogren's syndrome (often characterized, e.g., by multiple tissue antibodies, and/or a specific nonhistone ANA (SS-B)), diabetes mellitus (often characterized, e.g., by cell-mediated and humoral islet cell antibodies), and adrenergic drug resistance (including adrenergic drug resistance with asthma or cystic fibrosis) (often characterized, e.g., by beta-adrenergic receptor antibodies).

Additional disorders that may have an autoimmune component that may be treated, prevented, diagnosed and/or prognosed with the compositions of the invention include, but are not limited to, chronic active hepatitis (often characterized, e.g., by smooth muscle antibodies), primary biliary cirrhosis (often characterized, e.g., by mitochondria antibodies), other endocrine gland failure (often characterized, e.g., by specific tissue antibodies in some cases), vitiligo (often characterized, e.g., by melanocyte antibodies), vasculitis (often characterized, e.g., by Ig and complement in vessel walls and/or low serum complement), post-MI (often characterized, e.g., by myocardial antibodies), cardiotomy syndrome (often characterized, e.g., by myocardial antibodies), urticaria (often characterized, e.g., by IgG and IgM antibodies to IgE), atopic dermatitis (often characterized, e.g., by IgG and IgM antibodies to IgE), asthma (often characterized, e.g., by IgG and IgM antibodies to IgE), and many other inflammatory, granulomatous, degenerative, and atrophic disorders.

In a preferred embodiment, the autoimmune diseases and disorders and/or conditions associated with the diseases and disorders recited above are treated, prevented, diagnosed and/or prognosed using for example, antagonists or agonists, polypeptides or polynucleotides, or antibodies of the present invention. In a specific preferred embodiment, rheumatoid arthritis is treated, prevented, and/or diagnosed using polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention.

In another specific preferred embodiment, systemic lupus erythematosus is treated, prevented, and/or diagnosed using polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention. In another specific preferred embodiment, idiopathic thrombocytopenia purpura is treated, prevented, and/or diagnosed using polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention.

In another specific preferred embodiment IgA nephropathy is treated, prevented, and/or diagnosed using polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention.

In a preferred embodiment, the autoimmune diseases and disorders and/or conditions associated with the diseases and disorders recited above are treated, prevented, diagnosed and/or prognosed using polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention

In preferred embodiments, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a immunosuppressive agent(s).

Polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, prognosing, and/or diagnosing diseases, disorders, and/or conditions of hematopoietic cells. Polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention could be used to increase differentiation and proliferation of hematopoietic cells, including the pluripotent stem cells, in an effort to treat or prevent those diseases, disorders, and/or conditions associated with a decrease in certain (or many) types hematopoietic cells, including but not limited to, leukopenia, neutropenia, anemia, and thrombocytopenia. Alternatively, Polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention could be used to increase differentiation and proliferation of hematopoietic cells, including the pluripotent stem cells, in an effort to treat or prevent those diseases, disorders, and/or conditions associated with an increase in certain (or many) types of hematopoietic cells, including but not limited to, histiocytosis.

Allergic reactions and conditions, such as asthma (particularly allergic asthma) or other respiratory problems, may also be treated, prevented, diagnosed and/or prognosed using polypeptides, antibodies, or polynucleotides of the invention, and/or agonists or antagonists thereof. Moreover, these molecules can be used to treat, prevent, prognose, and/or diagnose anaphylaxis, hypersensitivity to an antigenic molecule, or blood group incompatibility.

Additionally, polypeptides or polynucleotides of the invention, and/or agonists or antagonists thereof, may be used to treat, prevent, diagnose and/or prognose IgE-mediated allergic reactions. Such allergic reactions include, but are not limited to, asthma, rhinitis, and eczema. In specific embodiments, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to modulate IgE concentrations in vitro or in vivo.

Moreover, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention have uses in the diagnosis, prognosis, prevention, and/or treatment of inflammatory conditions. For example, since polypeptides, antibodies, or polynucleotides of the invention, and/or agonists or antagonists of the invention may inhibit the activation, proliferation and/or differentiation of cells involved in an inflammatory response, these molecules can be used to prevent and/or treat chronic and acute inflammatory conditions. Such inflammatory conditions include, but are not limited to, for example, inflammation associated with infection (e.g., septic shock, sepsis, or systemic inflammatory response syndrome), ischemia-reperfusion injury, endotoxin lethality, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine induced lung injury, inflammatory bowel disease, Crohn's disease, over production of cytokines (e.g., TNF or IL-1.), respiratory disorders (e.g., asthma and allergy); gastrointestinal disorders (e.g., inflammatory bowel disease); cancers (e.g., gastric, ovarian, lung, bladder, liver, and breast); CNS disorders (e.g., multiple sclerosis; ischemic brain injury and/or stroke, traumatic brain injury, neurodegenerative disorders (e.g., Parkinson's disease and Alzheimer's disease); AIDS-related dementia; and prion disease); cardiovascular disorders (e.g., atherosclerosis, myocarditis, cardiovascular disease, and cardiopulmonary bypass complications); as well as many additional diseases, conditions, and disorders that are characterized by inflammation (e.g., hepatitis, rheumatoid arthritis, gout, trauma, pancreatitis, sarcoidosis, dermatitis, renal ischemia-reperfusion injury, Grave's disease, systemic lupus erythematosus, diabetes mellitus, and allogenic transplant rejection).

Because inflammation is a fundamental defense mechanism, inflammatory disorders can effect virtually any tissue of the body. Accordingly, polynucleotides, polypeptides, and antibodies of the invention, as well as agonists or antagonists thereof, have uses in the treatment of tissue-specific inflammatory disorders, including, but not limited to, adrenalitis, alveolitis, angiocholecystitis, appendicitis, balanitis, blepharitis, bronchitis, bursitis, carditis, cellulitis, cervicitis, cholecystitis, chorditis, cochlitis, colitis, conjunctivitis, cystitis, dermatitis, diverticulitis, encephalitis, endocarditis, esophagitis, eustachitis, fibrositis, folliculitis, gastritis, gastroenteritis, gingivitis, glossitis, hepatosplenitis, keratitis, labyrinthitis, laryngitis, lymphangitis, mastitis, media otitis, meningitis, metritis, mucitis, myocarditis, myosititis, myringitis, nephritis, neuritis, orchitis, osteochondritis, otitis, pericarditis, peritendonitis, peritonitis, pharyngitis, phlebitis, poliomyelitis, prostatitis, pulpitis, retinitis, rhinitis, salpingitis, scieritis, scierochoroiditis, scrotitis, sinusitis, spondylitis, steatitis, stomatitis, synovitis, syringitis, tendonitis, tonsillitis, urethritis, and vaginitis.

In specific embodiments, polypeptides, antibodies, or polynucleotides of the invention, and/or agonists or antagonists thereof, are useful to diagnose, prognose, prevent, and/or treat organ transplant rejections and graft-versus-host disease. Organ rejection occurs by host immune cell destruction of the transplanted tissue through an immune response. Similarly, an immune response is also involved in GVHD, but, in this case, the foreign transplanted immune cells destroy the host tissues. Polypeptides, antibodies, or polynucleotides of the invention, and/or agonists or antagonists thereof, that inhibit an immune response, particularly the activation, proliferation, differentiation, or chemotaxis of T-cells, may be an effective therapy in preventing organ rejection or GVHD. In specific embodiments, polypeptides, antibodies, or polynucleotides of the invention, and/or agonists or antagonists thereof, that inhibit an immune response, particularly the activation, proliferation, differentiation, or chemotaxis of T-cells, may be an effective therapy in preventing experimental allergic and hyperacute xenograft rejection.

In other embodiments, polypeptides, antibodies, or polynucleotides of the invention, and/or agonists or antagonists thereof, are useful to diagnose, prognose, prevent, and/or treat immune complex diseases, including, but not limited to, serum sickness, post streptococcal glomerulonephritis, polyarteritis nodosa, and immune complex-induced vasculitis.

Polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the invention can be used to treat, detect, and/or prevent infectious agents. For example, by increasing the immune response, particularly increasing the proliferation activation and/or differentiation of B and/or T cells, infectious diseases may be treated, detected, and/or prevented. The immune response may be increased by either enhancing an existing immune response, or by initiating a new immune response. Alternatively, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may also directly inhibit the infectious agent (refer to section of application listing infectious agents, etc), without necessarily eliciting an immune response.

In another embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a vaccine adjuvant that enhances immune responsiveness to an antigen. In a specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an adjuvant to enhance tumor-specific immune responses.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an adjuvant to enhance anti-viral immune responses. Anti-viral immune responses that may be enhanced using the compositions of the invention as an adjuvant, include virus and virus associated diseases or symptoms described herein or otherwise known in the art. In specific embodiments, the compositions of the invention are used as an adjuvant to enhance an immune response to a virus, disease, or symptom selected from the group consisting of: AIDS, meningitis, Dengue, EBV, and hepatitis (e.g., hepatitis B). In another specific embodiment, the compositions of the invention are used as an adjuvant to enhance an immune response to a virus, disease, or symptom selected from the group consisting of: HIV/AIDS, respiratory syncytial virus, Dengue, rotavirus, Japanese B encephalitis, influenza A and B, parainfluenza, measles, cytomegalovirus, rabies, Junin, Chikungunya, Rift Valley Fever, herpes simplex, and yellow fever.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an adjuvant to enhance anti-bacterial or anti-fungal immune responses. Anti-bacterial or anti-fungal immune responses that may be enhanced using the compositions of the invention as an adjuvant, include bacteria or fungus and bacteria or fungus associated diseases or symptoms described herein or otherwise known in the art. In specific embodiments, the compositions of the invention are used as an adjuvant to enhance an immune response to a bacteria or fungus, disease, or symptom selected from the group consisting of: tetanus, Diphtheria, botulism, and meningitis type B.

In another specific embodiment, the compositions of the invention are used as an adjuvant to enhance an immune response to a bacteria or fungus, disease, or symptom selected from the group consisting of: Vibrio cholerae, Mycobacterium leprae, Salmonella typhi, Salmonella paratyphi, Meisseria meningitidis, Streptococcus pneumoniae, Group B streptococcus, Shigella spp., Enterotoxigenic Escherichia coli, Enterohemorrhagic E. coli, and Borrelia burgdorferi.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an adjuvant to enhance anti-parasitic immune responses. Anti-parasitic immune responses that may be enhanced using the compositions of the invention as an adjuvant, include parasite and parasite associated diseases or symptoms described herein or otherwise known in the art. In specific embodiments, the compositions of the invention are used as an adjuvant to enhance an immune response to a parasite. In another specific embodiment, the compositions of the invention are used as an adjuvant to enhance an immune response to Plasmodium (malaria) or Leishmania.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention may also be employed to treat infectious diseases including silicosis, sarcoidosis, and idiopathic pulmonary fibrosis; for example, by preventing the recruitment and activation of mononuclear phagocytes.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an antigen for the generation of antibodies to inhibit or enhance immune mediated responses against polypeptides of the invention.

In one embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are administered to an animal (e.g., mouse, rat, rabbit, hamster, guinea pig, pigs, micro-pig, chicken, camel, goat, horse, cow, sheep, dog, cat, non-human primate, and human, most preferably human) to boost the immune system to produce increased quantities of one or more antibodies (e.g., IgG, IgA, IgM, and IgE), to induce higher affinity antibody production and immunoglobulin class switching (e.g., IgG, IgA, IgM, and IgE), and/or to increase an immune response.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a stimulator of B cell responsiveness to pathogens.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an activator of T cells.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent that elevates the immune status of an individual prior to their receipt of immunosuppressive therapies.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent to induce higher affinity antibodies.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent to increase serum immunoglobulin concentrations.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent to accelerate recovery of immunocompromised individuals.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent to boost immunoresponsiveness among aged populations and/or neonates.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an immune system enhancer prior to, during, or after bone marrow transplant and/or other transplants (e.g., allogeneic or xenogeneic organ transplantation). With respect to transplantation, compositions of the invention may be administered prior to, concomitant with, and/or after transplantation. In a specific embodiment, compositions of the invention are administered after transplantation, prior to the beginning of recovery of T-cell populations. In another specific embodiment, compositions of the invention are first administered after transplantation after the beginning of recovery of T cell populations, but prior to full recovery of B cell populations.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent to boost immunoresponsiveness among individuals having an acquired loss of B cell function. Conditions resulting in an acquired loss of B cell function that may be ameliorated or treated by administering the polypeptides, antibodies, polynucleotides and/or agonists or antagonists thereof, include, but are not limited to, HIV Infection, AIDS, bone marrow transplant, and B cell chronic lymphocytic leukemia (CLL).

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent to boost immunoresponsiveness among individuals having a temporary immune deficiency. Conditions resulting in a temporary immune deficiency that may be ameliorated or treated by administering the polypeptides, antibodies, polynucleotides and/or agonists or antagonists thereof, include, but are not limited to, recovery from viral infections (e.g., influenza), conditions associated with malnutrition, recovery from infectious mononucleosis, or conditions associated with stress, recovery from measles, recovery from blood transfusion, and recovery from surgery.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a regulator of antigen presentation by monocytes, dendritic cells, and/or B-cells. In one embodiment, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention enhance antigen presentation or antagonizes antigen presentation in vitro or in vivo. Moreover, in related embodiments, said enhancement or antagonism of antigen presentation may be useful as an anti-tumor treatment or to modulate the immune system.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as an agent to direct an individual's immune system towards development of a humoral response (i.e. TH2) as opposed to a TH1 cellular response.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a means to induce tumor proliferation and thus make it more susceptible to anti-neoplastic agents. For example, multiple myeloma is a slowly dividing disease and is thus refractory to virtually all anti-neoplastic regimens. If these cells were forced to proliferate more rapidly their susceptibility profile would likely change.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a stimulator of B cell production in pathologies such as AIDS, chronic lymphocyte disorder and/or Common Variable Immunodificiency.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a therapy for generation and/or regeneration of lymphoid tissues following surgery, trauma or genetic defect. In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used in the pretreatment of bone marrow samples prior to transplant.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a gene-based therapy for genetically inherited disorders resulting in immuno-incompetence/immunodeficiency such as observed among SCID patients.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a means of activating monocytes/macrophages to defend against parasitic diseases that effect monocytes such as Leishmania.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a means of regulating secreted cytokines that are elicited by polypeptides of the invention.

In another embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used in one or more of the applications described herein, as they may apply to veterinary medicine.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a means of blocking various aspects of immune responses to foreign agents or self. Examples of diseases or conditions in which blocking of certain aspects of immune responses may be desired include autoimmune disorders such as lupus, and arthritis, as well as immunoresponsiveness to skin allergies, inflammation, bowel disease, injury and diseases/disorders associated with pathogens.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a therapy for preventing the B cell proliferation and Ig secretion associated with autoimmune diseases such as idiopathic thrombocytopenic purpura, systemic lupus erythematosus and multiple sclerosis.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a inhibitor of B and/or T cell migration in endothelial cells. This activity disrupts tissue architecture or cognate responses and is useful, for example in disrupting immune responses, and blocking sepsis.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a therapy for chronic hypergammaglobulinemia evident in such diseases as monoclonal gammopathy of undetermined significance (MGUS), Waldenstrom's disease, related idiopathic monoclonal gammopathies, and plasmacytomas.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention may be employed for instance to inhibit polypeptide chemotaxis and activation of macrophages and their precursors, and of neutrophils, basophils, B lymphocytes and some T-cell subsets, e.g., activated and CD8 cytotoxic T cells and natural killer cells, in certain autoimmune and chronic inflammatory and infective diseases. Examples of autoimmune diseases are described herein and include multiple sclerosis, and insulin-dependent diabetes.

The polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention may also be employed to treat idiopathic hyper-eosinophilic syndrome by, for example, preventing eosinophil production and migration.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used to enhance or inhibit complement mediated cell lysis.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used to enhance or inhibit antibody dependent cellular cytotoxicity.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention may also be employed for treating atherosclerosis, for example, by preventing monocyte infiltration in the artery wall.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention may be employed to treat adult respiratory distress syndrome (ARDS).

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention may be useful for stimulating wound and tissue repair, stimulating angiogenesis, and/or stimulating the repair of vascular or lymphatic diseases or disorders. Additionally, agonists and antagonists of the invention may be used to stimulate the regeneration of mucosal surfaces.

In a specific embodiment, polynucleotides or polypeptides, and/or agonists thereof are used to diagnose, prognose, treat, and/or prevent a disorder characterized by primary or acquired immunodeficiency, deficient serum immunoglobulin production, recurrent infections, and/or immune system dysfunction. Moreover, polynucleotides or polypeptides, and/or agonists thereof may be used to treat or prevent infections of the joints, bones, skin, and/or parotid glands, blood-borne infections (e.g., sepsis, meningitis, septic arthritis, and/or osteomyelitis), autoimmune diseases (e.g., those disclosed herein), inflammatory disorders, and malignancies, and/or any disease or disorder or condition associated with these infections, diseases, disorders and/or malignancies) including, but not limited to, CVID, other primary immune deficiencies, HIV disease, CLL, recurrent bronchitis, sinusitis, otitis media, conjunctivitis, pneumonia, hepatitis, meningitis, herpes zoster (e.g., severe herpes zoster), and/or pneumocystis carnii. Other diseases and disorders that may be prevented, diagnosed, prognosed, and/or treated with polynucleotides or polypeptides, and/or agonists of the present invention include, but are not limited to, HIV infection, HTLV-BLV infection, lymphopenia, phagocyte bactericidal dysfunction anemia, thrombocytopenia, and hemoglobinuria.

In another embodiment, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention are used to treat, and/or diagnose an individual having common variable immunodeficiency disease (“CVID”; also known as “acquired agammaglobulinemia” and “acquired hypogammaglobulinemia”) or a subset of this disease.

In a specific embodiment, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to diagnose, prognose, prevent, and/or treat cancers or neoplasms including immune cell or immune tissue-related cancers or neoplasms. Examples of cancers or neoplasms that may be prevented, diagnosed, or treated by polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention include, but are not limited to, acute myelogenous leukemia, chronic myelogenous leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphocytic anemia (ALL) Chronic lymphocyte leukemia, plasmacytomas, multiple myeloma, Burkitt's lymphoma, EBV-transformed diseases, and/or diseases and disorders described in the section entitled “Hyperproliferative Disorders” elsewhere herein.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a therapy for decreasing cellular proliferation of Large B-cell Lymphomas.

In another specific embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are used as a means of decreasing the involvement of B cells and Ig associated with Chronic Myelogenous Leukemia.

In specific embodiments, the compositions of the invention are used as an agent to boost immunoresponsiveness among B cell immunodeficient individuals, such as, for example, an individual who has undergone a partial or complete splenectomy.

Antagonists of the invention include, for example, binding and/or inhibitory antibodies, antisense nucleic acids, ribozymes or soluble forms of the polypeptides of the present invention (e.g., Fc fusion protein; see, e.g., Example 9). Agonists of the invention include, for example, binding or stimulatory antibodies, and soluble forms of the polypeptides (e.g., Fc fusion proteins; see, e.g., Example 9). polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention may be employed in a composition with a pharmaceutically acceptable carrier, e.g., as described herein.

In another embodiment, polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention are administered to an animal (including, but not limited to, those listed above, and also including transgenic animals) incapable of producing functional endogenous antibody molecules or having an otherwise compromised endogenous immune system, but which is capable of producing human immunoglobulin molecules by means of a reconstituted or partially reconstituted immune system from another animal (see, e.g., published PCT Application Nos. WO98/24893, WO/9634096, WO/9633735, and WO/9110741). Administration of polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention to such animals is useful for the generation of monoclonal antibodies against the polypeptides, antibodies, polynucleotides and/or agonists or antagonists of the present invention.

Blood-Related Disorders

The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to modulate hemostatic (the stopping of bleeding) or thrombolytic (clot dissolving) activity. For example, by increasing hemostatic or thrombolytic activity, polynucleotides or polypeptides, and/or agonists or antagonists of the present invention could be used to treat or prevent blood coagulation diseases, disorders, and/or conditions (e.g., afibrinogenemia, factor deficiencies, hemophilia), blood platelet diseases, disorders, and/or conditions (e.g., thrombocytopenia), or wounds resulting from trauma, surgery, or other causes. Alternatively, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention that can decrease hemostatic or thrombolytic activity could be used to inhibit or dissolve clotting. These molecules could be important in the treatment or prevention of heart attacks (infarction), strokes, or scarring.

In specific embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to prevent, diagnose, prognose, and/or treat thrombosis, arterial thrombosis, venous thrombosis, thromboembolism, pulmonary embolism, atherosclerosis, myocardial infarction, transient ischemic attack, unstable angina. In specific embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used for the prevention of occlusion of saphenous grafts, for reducing the risk of periprocedural thrombosis as might accompany angioplasty procedures, for reducing the risk of stroke in patients with atrial fibrillation including nonrheumatic atrial fibrillation, for reducing the risk of embolism associated with mechanical heart valves and or mitral valves disease. Other uses for the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention, include, but are not limited to, the prevention of occlusions in extrcorporeal devices (e.g., intravascular canulas, vascular access shunts in hemodialysis patients, hemodialysis machines, and cardiopulmonary bypass machines).

In another embodiment, a polypeptide of the invention, or polynucleotides, antibodies, agonists, or antagonists corresponding to that polypeptide, may be used to prevent, diagnose, prognose, and/or treat diseases and disorders of the blood and/or blood forming organs associated with the tissue(s) in which the polypeptide of the invention is expressed, including one, two, three, four, five, or more tissues disclosed in Table 1B.2, column 5 (Tissue Distribution Library Code).

The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to modulate hematopoietic activity (the formation of blood cells). For example, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to increase the quantity of all or subsets of blood cells, such as, for example, erythrocytes, lymphocytes (B or T cells), myeloid cells (e.g., basophils, eosinophils, neutrophils, mast cells, macrophages) and platelets. The ability to decrease the quantity of blood cells or subsets of blood cells may be useful in the prevention, detection, diagnosis and/or treatment of anemias and leukopenias described below. Alternatively, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to decrease the quantity of all or subsets of blood cells, such as, for example, erythrocytes, lymphocytes (B or T cells), myeloid cells (e.g., basophils, eosinophils, neutrophils, mast cells, macrophages) and platelets. The ability to decrease the quantity of blood cells or subsets of blood cells may be useful in the prevention, detection, diagnosis and/or treatment of leukocytoses, such as, for example eosinophilia.

The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be used to prevent, treat, or diagnose blood dyscrasia.

Anemias are conditions in which the number of red blood cells or amount of hemoglobin (the protein that carries oxygen) in them is below normal. Anemia may be caused by excessive bleeding, decreased red blood cell production, or increased red blood cell destruction (hemolysis). The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, and/or diagnosing anemias. Anemias that may be treated prevented or diagnosed by the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention include iron deficiency anemia, hypochromic anemia, microcytic anemia, chlorosis, hereditary sideroblastic anemia, idiopathic acquired sideroblastic anemia, red cell aplasia, megaloblastic anemia (e.g., pernicious anemia, (vitamin B12 deficiency) and folic acid deficiency anemia), aplastic anemia, hemolytic anemias (e.g., autoimmune helolytic anemia, microangiopathic hemolytic anemia, and paroxysmal nocturnal hemoglobinuria). The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, and/or diagnosing anemias associated with diseases including but not limited to, anemias associated with systemic lupus erythematosus, cancers, lymphomas, chronic renal disease, and enlarged spleens. The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, and/or diagnosing anemias arising from drug treatments such as anemias associated with methyldopa, dapsonc, and/or sulfadrugs. Additionally, rhe polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, and/or diagnosing anemias associated with abnormal red blood cell architecture including but not limited to, hereditary spherocytosis, hereditary elliptocytosis, glucose-6-phosphate dehydrogenase deficiency, and sickle cell anemia.

The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, and/or diagnosing hemoglobin abnormalities, (e.g., those associated with sickle cell anemia, hemoglobin C disease, hemoglobin S-C disease, and hemoglobin E disease). Additionally, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating thalassemias, including, but not limited to major and minor forms of alpha-thalassemia and beta-thalassemia.

In another embodiment, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating bleeding disorders including, but not limited to, thrombocytopenia (e.g., idiopathic thrombocytopenic purpura, and thrombotic thrombocytopenic purpura), Von Willebrand's disease, hereditary platelet disorders (e.g., storage pool disease such as Chediak-Higashi and Hermansky-Pudlak syndromes, thromboxane A2 dysfunction, thromboasthenia, and Bernard-Soulier syndrome), hemolytic-uremic syndrome, hemophelias such as hemophelia A or Factor VII deficiency and Christmas disease or Factor IX deficiency, Hereditary Hemorhhagic Telangiectsia, also known as Rendu-Osler-Weber syndrome, allergic purpura (Henoch Schonlein purpura) and disseminated intravascular coagulation.

The effect of the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention on the clotting time of blood may be monitored using any of the clotting tests known in the art including, but not limited to, whole blood partial thromboplastin time (PTT), the activated partial thromboplastin time (aPTT), the activated clotting time (ACT), the recalcified activated clotting time, or the Lee-White Clotting time.

Several diseases and a variety of drugs can cause platelet dysfunction. Thus, in a specific embodiment, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating acquired platelet dysfunction such as platelet dysfunction accompanying kidney failure, leukemia, multiple myeloma, cirrhosis of the liver, and systemic lupus erythematosus as well as platelet dysfunction associated with drug treatments, including treatment with aspirin, ticlopidine, nonsteroidal anti-inflammatory drugs (used for arthritis, pain, and sprains), and penicillin in high doses.

In another embodiment, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating diseases and disorders characterized by or associated with increased or decreased numbers of white blood cells. Leukopenia occurs when the number of white blood cells decreases below normal. Leukopenias include, but are not limited to, neutropenia and lymphocytopenia. An increase in the number of white blood cells compared to normal is known as leukocytosis. The body generates increased numbers of white blood cells during infection. Thus, leukocytosis may simply be a normal physiological parameter that reflects infection. Alternatively, leukocytosis may be an indicator of injury or other disease such as cancer. Leokocytoses, include but are not limited to, eosinophilia, and accumulations of macrophages. In specific embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating leukopenia. In other specific embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating leukocytosis.

Leukopenia may be a generalized decreased in all types of white blood cells, or may be a specific depletion of particular types of white blood cells. Thus, in specific embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating decreases in neutrophil numbers, known as neutropenia. Neutropenias that may be diagnosed, prognosed, prevented, and/or treated by the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention include, but are not limited to, infantile genetic agranulocytosis, familial neutropenia, cyclic neutropenia, neutropenias resulting from or associated with dietary deficiencies (e.g., vitamin B 12 deficiency or folic acid deficiency), neutropenias resulting from or associated with drug treatments (e.g., antibiotic regimens such as penicillin treatment, sulfonamide treatment, anticoagulant treatment, anticonvulsant drugs, anti-thyroid drugs, and cancer chemotherapy), and neutropenias resulting from increased neutrophil destruction that may occur in association with some bacterial or viral infections, allergic disorders, autoimmune diseases, conditions in which an individual has an enlarged spleen (e.g., Felty syndrome, malaria and sarcoidosis), and some drug treatment regimens.

The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating lymphocytopenias (decreased numbers of B and/or T lymphocytes), including, but not limited lymphocytopenias resulting from or associated with stress, drug treatments (e.g., drug treatment with corticosteroids, cancer chemotherapies, and/or radiation therapies), AIDS infection and/or other diseases such as, for example, cancer, rheumatoid arthritis, systemic lupus erythematosus, chronic infections, some viral infections and/or hereditary disorders (e.g., DiGeorge syndrome, Wiskott-Aldrich Syndome, severe combined immunodeficiency, ataxia telangiectsia).

The polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating diseases and disorders associated with macrophage numbers and/or macrophage function including, but not limited to, Gaucher's disease, Niemann-Pick disease, Letterer-Siwe disease and Hand-Schuller-Christian disease.

In another embodiment, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating diseases and disorders associated with eosinophil numbers and/or eosinophil function including, but not limited to, idiopathic hypereosinophilic syndrome, eosinophilia-myalgia syndrome, and Hand-Schuller-Christian disease.

In yet another embodiment, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating leukemias and lymphomas including, but not limited to, acute lymphocytic (lymphpblastic) leukemia (ALL), acute myeloid (myelocytic, myelogenous, myeloblastic, or myelomonocytic) leukemia, chronic lymphocytic leukemia (e.g., B cell leukemias, T cell leukemias, Sezary syndrome, and Hairy cell leukenia), chronic myelocytic (myeloid, myelogenous, or granulocytic) leukemia, Hodgkin's lymphoma, non-hodgkin's lymphoma, Burkitt's lymphoma, and mycosis fungoides.

In other embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in diagnosing, prognosing, preventing, and/or treating diseases and disorders of plasma cells including, but not limited to, plasma cell dyscrasias, monoclonal gammaopathies, monoclonal gammopathies of undetermined significance, multiple myeloma, macroglobulinemia, Waldenstrom's macroglobulinemia, cryoglobulinemia, and Raynaud's phenomenon.

In other embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in treating, preventing, and/or diagnosing myeloproliferative disorders, including but not limited to, polycythemia vera, relative polycythemia, secondary polycythemia, myelofibrosis, acute myelofibrosis, agnogenic myelod metaplasia, thrombocythemia, (including both primary and secondary thrombocythemia) and chronic myelocytic leukemia.

In other embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful as a treatment prior to surgery, to increase blood cell production.

In other embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful as an agent to enhance the migration, phagocytosis, superoxide production, antibody dependent cellular cytotoxicity of neutrophils, eosionophils and macrophages.

In other embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful as an agent to increase the number of stem cells in circulation prior to stem cells pheresis. In another specific embodiment, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful as an agent to increase the number of stem cells in circulation prior to platelet pheresis.

In other embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful as an agent to increase cytokine production.

In other embodiments, the polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention may be useful in preventing, diagnosing, and/or treating primary hematopoietic disorders.

Hyperproliferative Disorders

In certain embodiments, polynucleotides or polypeptides, or agonists or antagonists of the present invention can be used to treat or detect hyperproliferative disorders, including neoplasms. Polynucleotides or polypeptides, or agonists or antagonists of the present invention may inhibit the proliferation of the disorder through direct or indirect interactions. Alternatively, Polynucleotides or polypeptides, or agonists or antagonists of the present invention may proliferate other cells which can inhibit the hyperproliferative disorder.

For example, by increasing an immune response, particularly increasing antigenic qualities of the hyperproliferative disorder or by proliferating, differentiating, or mobilizing T-cells, hyperproliferative disorders can be treated. This immune response may be increased by either enhancing an existing immune response, or by initiating a new immune response. Alternatively, decreasing an immune response may also be a method of treating hyperproliferative disorders, such as a chemotherapeutic agent.

Examples of hyperproliferative disorders that can be treated or detected by polynucleotides or polypeptides, or agonists or antagonists of the present invention include, but are not limited to neoplasms located in the: colon, abdomen, bone, breast, digestive system, liver, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid), eye, head and neck, nervous (central and peripheral), lymphatic system, pelvis, skin, soft tissue, spleen, thorax, and urogenital tract.

Similarly, other hyperproliferative disorders can also be treated or detected by polynucleotides or polypeptides, or agonists or antagonists of the present invention. Examples of such hyperproliferative disorders include, but are not limited to: Acute Childhood Lymphoblastic Leukemia, Acute Lymphoblastic Leukemia, Acute Lymphocytic Leukemia, Acute Myeloid Leukemia, Adrenocortical Carcinoma, Adult (Primary) Hepatocellular Cancer, Adult (Primary) Liver Cancer, Adult Acute Lymphocytic Leukemia, Adult Acute Myeloid Leukemia, Adult Hodgkin's Disease, Adult Hodgkin's Lymphoma, Adult Lymphocytic Leukemia, Adult Non-Hodgkin's Lymphoma, Adult Primary Liver Cancer, Adult Soft Tissue Sarcoma, AIDS-Related Lymphoma, AIDS-Related Malignancies, Anal Cancer, Astrocytoma, Bile Duct Cancer, Bladder Cancer, Bone Cancer, Brain Stem Glioma, Brain Tumors, Breast Cancer, Cancer of the Renal Pelvis and Ureter, Central Nervous System (Primary) Lymphoma, Central Nervous System Lymphoma, Cerebellar Astrocytoma, Cerebral Astrocytoma, Cervical Cancer, Childhood (Primary) Hepatocellular Cancer, Childhood (Primary) Liver Cancer, Childhood Acute Lymphoblastic Leukemia, Childhood Acute Myeloid Leukemia, Childhood Brain Stem Glioma, Childhood Cerebellar Astrocytoma, Childhood Cerebral Astrocytoma, Childhood Extracranial Germ Cell Tumors, Childhood Hodgkin's Disease, Childhood Hodgkin's Lymphoma, Childhood Hypothalamic and Visual Pathway Glioma, Childhood Lymphoblastic Leukemia, Childhood Medulloblastoma, Childhood Non-Hodgkin's Lymphoma, Childhood Pineal and Supratentorial Primitive Neuroectodermal Tumors, Childhood Primary Liver Cancer, Childhood Rhabdomyosarcoma, Childhood Soft Tissue Sarcoma, Childhood Visual Pathway and Hypothalamic Glioma, Chronic Lymphocytic Leukemia, Chronic Myelogenous Leukemia, Colon Cancer, Cutaneous T-Cell Lymphoma, Endocrine Pancreas Islet Cell Carcinoma, Endometrial Cancer, Ependymoma, Epithelial Cancer, Esophageal Cancer, Ewing's Sarcoma and Related Tumors, Exocrine Pancreatic Cancer, Extracranial Germ Cell Tumor, Extragonadal Germ Cell Tumor, Extrahepatic Bile Duct Cancer, Eye Cancer, Female Breast Cancer, Gaucher's Disease, Gallbladder Cancer, Gastric Cancer, Gastrointestinal Carcinoid Tumor, Gastrointestinal Tumors, Germ Cell Tumors, Gestational Trophoblastic Tumor, Hairy Cell Leukemia, Head and Neck Cancer, Hepatocellular Cancer, Hodgkin's Disease, Hodgkin's Lymphoma, Hypergammaglobulinemia, Hypopharyngeal Cancer, Intestinal Cancers, Intraocular Melanoma, Islet Cell Carcinoma, Islet Cell Pancreatic Cancer, Kaposi's Sarcoma, Kidney Cancer, Laryngeal Cancer, Lip and Oral Cavity Cancer, Liver Cancer, Lung Cancer, Lymphoproliferative Disorders, Macroglobulinemia, Male Breast Cancer, Malignant Mesothelioma, Malignant Thymoma, Medulloblastoma, Melanoma, Mesothelioma, Metastatic Occult Primary Squamous Neck Cancer, Metastatic Primary Squamous Neck Cancer, Metastatic Squamous Neck Cancer, Multiple Myeloma, Multiple Myeloma/Plasma Cell Neoplasm, Myelodysplastic Syndrome, Myelogenous Leukemia, Myeloid Leukemia, Myeloproliferative Disorders, Nasal Cavity and Paranasal Sinus Cancer, Nasopharyngeal Cancer, Neuroblastoma, Non-Hodgkin's Lymphoma During Pregnancy, Nonmelanoma Skin Cancer, Non-Small Cell Lung Cancer, Occult Primary Metastatic Squamous Neck Cancer, Oropharyngeal Cancer, Osteo/Malignant Fibrous Sarcoma, Osteosarcoma/Malignant Fibrous Histiocytoma, Osteosarcomal/Malignant Fibrous Histiocytoma of Bone, Ovarian Epithelial Cancer, Ovarian Germ Cell Tumor, Ovarian Low Malignant Potential Tumor, Pancreatic Cancer, Paraproteinemias, Purpura, Parathyroid Cancer, Penile Cancer, Pheochromocytoma, Pituitary Tumor, Plasma Cell Neoplasm/Multiple Myeloma, Primary Central Nervous System Lymphoma, Primary Liver Cancer, Prostate Cancer, Rectal Cancer, Renal Cell Cancer, Renal Pelvis and Ureter Cancer, Retinoblastoma, Rhabdomyosarcoma, Salivary Gland Cancer, Sarcoidosis Sarcomas, Sezary Syndrome, Skin Cancer, Small Cell Lung Cancer, Small Intestine Cancer, Soft Tissue Sarcoma, Squamous Neck Cancer, Stomach Cancer, Supratentorial Primitive Neuroectodermal and Pineal Tumors, T-Cell Lymphoma, Testicular Cancer, Thymoma, Thyroid Cancer, Transitional Cell Cancer of the Renal Pelvis and Ureter, Transitional Renal Pelvis and Ureter Cancer, Trophoblastic Tumors, Ureter and Renal Pelvis Cell Cancer, Urethral Cancer, Uterine Cancer, Uterine Sarcoma, Vaginal Cancer, Visual Pathway and Hypothalamic Glioma, Vulvar Cancer, Waldenstrom's Macroglobulinemia, Wilms' Tumor, and any other hyperproliferative disease, besides neoplasia, located in an organ system listed above.

In another preferred embodiment, polynucleotides or polypeptides, or agonists or antagonists of the present invention are used to diagnose, prognose, prevent, and/or treat premalignant conditions and to prevent progression to a neoplastic or malignant state, including but not limited to those disorders described above. Such uses are indicated in conditions known or suspected of preceding progression to neoplasia or cancer, in particular, where non-neoplastic cell growth consisting of hyperplasia, metaplasia, or most particularly, dysplasia has occurred (for review of such abnormal growth conditions, see Robbins and Angell, 1976, Basic Pathology, 2d Ed., W. B. Saunders Co., Philadelphia, pp. 68-79.)

Hyperplasia is a form of controlled cell proliferation, involving an increase in cell number in a tissue or organ, without significant alteration in structure or function. Hyperplastic disorders which can be diagnosed, prognosed, prevented, and/or treated with compositions of the invention (including polynucleotides, polypeptides, agonists or antagonists) include, but are not limited to, angiofollicular mediastinal lymph node hyperplasia, angiolymphoid hyperplasia with eosinophilia, atypical melanocytic hyperplasia, basal cell hyperplasia, benign giant lymph node hyperplasia, cementum hyperplasia, congenital adrenal hyperplasia, congenital sebaceous hyperplasia, cystic hyperplasia, cystic hyperplasia of the breast, denture hyperplasia, ductal hyperplasia, endometrial hyperplasia, fibromuscular hyperplasia, focal epithelial hyperplasia, gingival hyperplasia, inflammatory fibrous hyperplasia, inflammatory papillary hyperplasia, intravascular papillary endothelial hyperplasia, nodular hyperplasia of prostate, nodular regenerative hyperplasia, pseudoepitheliomatous hyperplasia, senile sebaceous hyperplasia, and verrucous hyperplasia.

Metaplasia is a form of controlled cell growth in which one type of adult or fully differentiated cell substitutes for another type of adult cell. Metaplastic disorders which can be diagnosed, prognosed, prevented, and/or treated with compositions of the invention (including polynucleotides, polypeptides, agonists or antagonists) include, but are not limited to, agnogenic myeloid metaplasia, apocrine metaplasia, atypical metaplasia, autoparenchymatous metaplasia, connective tissue metaplasia, epithelial metaplasia, intestinal metaplasia, metaplastic anemia, metaplastic ossification, metaplastic polyps, myeloid metaplasia, primary myeloid metaplasia, secondary myeloid metaplasia, squamous metaplasia, squamous metaplasia of amnion, and symptomatic myeloid metaplasia.

Dysplasia is frequently a forerunner of cancer, and is found mainly in the epithelia; it is the most disorderly form of non-neoplastic cell growth, involving a loss in individual cell uniformity and in the architectural orientation of cells. Dysplastic cells often have abnormally large, deeply stained nuclei, and exhibit pleomorphism. Dysplasia characteristically occurs where there exists chronic irritation or inflammation. Dysplastic disorders which can be diagnosed, prognosed, prevented, and/or treated with compositions of the invention (including polynucleotides, polypeptides, agonists or antagonists) include, but are not limited to, anhidrotic ectodermal dysplasia, anterofacial dysplasia, asphyxiating thoracic dysplasia, atriodigital dysplasia, bronchopulmonary dysplasia, cerebral dysplasia, cervical dysplasia, chondroectodermal dysplasia, cleidocranial dysplasia, congenital ectodermal dysplasia, craniodiaphysial dysplasia, craniocarpotarsal dysplasia, craniometaphysial dysplasia, dentin dysplasia, diaphysial dysplasia, ectodermal dysplasia, enamel dysplasia, encephalo-ophthalmic dysplasia, dysplasia epiphysialis hemimelia, dysplasia epiphysialis multiplex, dysplasia epiphysialis punctata, epithelial dysplasia, faciodigitogenital dysplasia, familial fibrous dysplasia of jaws, familial white folded dysplasia, fibromuscular dysplasia, fibrous dysplasia of bone, florid osseous dysplasia, hereditary renal-retinal dysplasia, hidrotic ectodermal dysplasia, hypohidrotic ectodermal dysplasia, lymphopenic thymic dysplasia, mammary dysplasia, mandibulofacial dysplasia, metaphysial dysplasia, Mondini dysplasia, monostotic fibrous dysplasia, mucoepithelial dysplasia, multiple epiphysial dysplasia, oculoauriculovertebral dysplasia, oculodentodigital dysplasia, oculoveltebral dysplasia, odontogenic dysplasia, ophthalmomandibulomelic dysplasia, periapical cemental dysplasia, polyostotic fibrous dysplasia, pseudoachondroplastic spondyloepiphysial dysplasia, retinal dysplasia, septo-optic dysplasia, spondyloepiphysial dysplasia, and ventriculoradial dysplasia.

Additional pre-neoplastic disorders which can be diagnosed, prognosed, prevented, and/or treated with compositions of the invention (including polynucleotides, polypeptides, agonists or antagonists) include, but are not limited to, benign dysproliferative disorders (e.g., benign tumors, fibrocystic conditions, tissue hypertrophy, intestinal polyps, colon polyps, and esophageal dysplasia), leukoplakia, keratoses, Bowen's disease, Farmer's Skin, solar cheilitis, and solar keratosis.

In another embodiment, a polypeptide of the invention, or polynucleotides, antibodies, agonists, or antagonists corresponding to that polypeptide, may be used to diagnose and/or prognose disorders associated with the tissue(s) in which the polypeptide of the invention is expressed, including one, two, three, four, five, or more tissues disclosed in Table 1B.2, column 5 (Tissue Distribution Library Code).

In another embodiment, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention conjugated to a toxin or a radioactive isotope, as described herein, may be used to treat cancers and neoplasms, including, but not limited to those described herein. In a further preferred embodiment, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention conjugated to a toxin or a radioactive isotope, as described herein, may be used to treat acute myelogenous leukemia.

Additionally, polynucleotides, polypeptides, and/or agonists or antagonists of the invention may affect apoptosis, and therefore, would be useful in treating a number of diseases associated with increased cell survival or the inhibition of apoptosis. For example, diseases associated with increased cell survival or the inhibition of apoptosis that could be diagnosed, prognosed, prevented, and/or treated by polynucleotides, polypeptides, and/or agonists or antagonists of the invention, include cancers (such as follicular lymphomas, carcinomas with p53 mutations, and hormone-dependent tumors, including, but not limited to colon cancer, cardiac tumors, pancreatic cancer, melanoma, retinoblastoma, glioblastoma, lung cancer, intestinal cancer, testicular cancer, stomach cancer, neuroblastoma, myxoma, myoma, lymphoma, endothelioma, osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, adenoma, breast cancer, prostate cancer, Kaposi's sarcoma and ovarian cancer); autoimmune disorders such as, multiple sclerosis, Sjogren's syndrome, Hashimoto's thyroiditis, biliary cirrhosis, Behcet's disease, Crohn's disease, polymyositis, systemic lupus erythematosus and immune-related glomerulonephritis and rheumatoid arthritis) and viral infections (such as herpes viruses, pox viruses and adenoviruses), inflammation, graft v. host disease, acute graft rejection, and chronic graft rejection.

In preferred embodiments, polynucleotides, polypeptides, and/or agonists or antagonists of the invention are used to inhibit growth, progression, and/or metastasis of cancers, in particular those listed above.

Additional diseases or conditions associated with increased cell survival that could be diagnosed, prognosed, prevented, and/or treated by polynucleotides, polypeptides, and/or agonists or antagonists of the invention, include, but are not limited to, progression, and/or metastases of malignancies and related disorders such as leukemia (including acute leukemias (e.g., acute lymphocytic leukemia, acute myelocytic leukemia (including myeloblastic, promyelocytic, myelomonocytic, monocytic, and erythroleukemia)) and chronic leukemias (e.g., chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia)), polycythemia vera, lymphomas (e.g., Hodgkin's disease and non-Hodgkin's disease), multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, and solid tumors including, but not limited to, sarcomas and carcinomas such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, emangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma, and retinoblastoma.

Diseases associated with increased apoptosis that could be diagnosed, prognosed, prevented, and/or treated by polynucleotides, polypeptides, and/or agonists or antagonists of the invention, include AIDS; neurodegenerative disorders (such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, cerebellar degeneration and brain tumor or prior associated disease); autoimmune disorders (such as, multiple sclerosis, Sjogren's syndrome, Hashimoto's thyroiditis, biliary cirrhosis, Behcet's disease, Crohn's disease, polymyositis, systemic lupus erythematosus and immune-related glomerulonephritis and rheumatoid arthritis) myelodysplastic syndromes (such as aplastic anemia), graft v. host disease, ischemic injury (such as that caused by myocardial infarction, stroke and reperfusion injury), liver injury (e.g., hepatitis related liver injury, ischemia/reperfusion injury, cholestosis (bile duct injury) and liver cancer); toxin-induced liver disease (such as that caused by alcohol), septic shock, cachexia and anorexia.

Hyperproliferative diseases and/or disorders that could be diagnosed, prognosed, prevented, and/or treated by polynucleotides, polypeptides, and/or agonists or antagonists of the invention, include, but are not limited to, neoplasms located in the liver, abdomen, bone, breast, digestive system, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid), eye, head and neck, nervous system (central and peripheral), lymphatic system, pelvis, skin, soft tissue, spleen, thorax, and urogenital tract.

Similarly, other hyperproliferative disorders can also be diagnosed, prognosed, prevented, and/or treated by polynucleotides, polypeptides, and/or agonists or antagonists of the invention. Examples of such hyperproliferative disorders include, but are not limited to: hypergammaglobulinemia, lymphoproliferative disorders, paraproteinemias, purpura, sarcoidosis, Sezary Syndrome, Waldenstron's macroglobulinemia, Gaucher's Disease, histiocytosis, and any other hyperproliferative disease, besides neoplasia, located in an organ system listed above.

Another preferred embodiment utilizes polynucleotides of the present invention to inhibit aberrant cellular division, by gene therapy using the present invention, and/or protein fusions or fragments thereof.

Thus, the present invention provides a method for treating cell proliferative disorders by inserting into an abnormally proliferating cell a polynucleotide of the present invention, wherein said polynucleotide represses said expression.

Another embodiment of the present invention provides a method of treating cell-proliferative disorders in individuals comprising administration of one or more active gene copies of the present invention to an abnormally proliferating cell or cells. In a preferred embodiment, polynucleotides of the present invention is a DNA construct comprising a recombinant expression vector effective in expressing a DNA sequence encoding said polynucleotides. In another preferred embodiment of the present invention, the DNA construct encoding the polynucleotides of the present invention is inserted into cells to be treated utilizing a retrovirus, or more preferably an adenoviral vector (See G J. Nabel, et. al., PNAS 1999 96: 324-326, which is hereby incorporated by reference). In a most preferred embodiment, the viral vector is defective and will not transform non-proliferating cells, only proliferating cells. Moreover, in a preferred embodiment, the polynucleotides of the present invention inserted into proliferating cells either alone, or in combination with or fused to other polynucleotides, can then be modulated via an external stimulus (i.e. magnetic, specific small molecule, chemical, or drug administration, etc.), which acts upon the promoter upstream of said polynucleotides to induce expression of the encoded protein product. As such the beneficial therapeutic affect of the present invention may be expressly modulated (i.e. to increase, decrease, or inhibit expression of the present invention) based upon said external stimulus.

Polynucleotides of the present invention may be useful in repressing expression of oncogenic genes or antigens. By “repressing expression of the oncogenic genes” is intended the suppression of the transcription of the gene, the degradation of the gene transcript (pre-message RNA), the inhibition of splicing, the destruction of the messenger RNA, the prevention of the post-translational modifications of the protein, the destruction of the protein, or the inhibition of the normal function of the protein.

For local administration to abnormally proliferating cells, polynucleotides of the present invention may be administered by any method known to those of skill in the art including, but not limited to transfection, electroporation, microinjection of cells, or in vehicles such as liposomes, lipofectin, or as naked polynucleotides, or any other method described throughout the specification. The polynucleotide of the present invention may be delivered by known gene delivery systems such as, but not limited to, retroviral vectors (Gilboa, J. Virology 44:845 (1982); Hocke, Nature 320:275 (1986); Wilson, et al., Proc. Natl. Acad. Sci. U.S.A. 85:3014), vaccinia virus system (Chakrabarty et al., Mol. Cell Biol. 5:3403 (1985) or other efficient DNA delivery systems (Yates et al., Nature 313:812 (1985)) known to those skilled in the art. These references are exemplary only and are hereby incorporated by reference. In order to specifically deliver or transfect cells which are abnormally proliferating and spare non-dividing cells, it is preferable to utilize a retrovirus, or adenoviral (as described in the art and elsewhere herein) delivery system known to those of skill in the art. Since host DNA replication is required for retroviral DNA to integrate and the retrovirus will be unable to self replicate due to the lack of the retrovirus genes needed for its life cycle. Utilizing such a retroviral delivery system for polynucleotides of the present invention will target said gene and constructs to abnormally proliferating cells and will spare the non-dividing normal cells.

The polynucleotides of the present invention may be delivered directly to cell proliferative disorder/disease sites in internal organs, body cavities and the like by use of imaging devices used to guide an injecting needle directly to the disease site. The polynucleotides of the present invention may also be administered to disease sites at the time of surgical intervention.

By “cell proliferative disease” is meant any human or animal disease or disorder, affecting any one or any combination of organs, cavities, or body parts, which is characterized by single or multiple local abnormal proliferations of cells, groups of cells, or tissues, whether benign or malignant.

Any amount of the polynucleotides of the present invention may be administered as long as it has a biologically inhibiting effect on the proliferation of the treated cells. Moreover, it is possible to administer more than one of the polynucleotide of the present invention simultaneously to the same site. By “biologically inhibiting” is meant partial or total growth inhibition as well as decreases in the rate of proliferation or growth of the cells. The biologically inhibitory dose may be determined by assessing the effects of the polynucleotides of the present invention on target malignant or abnormally proliferating cell growth in tissue culture, tumor growth in animals and cell cultures, or any other method known to one of ordinary skill in the art.

The present invention is further directed to antibody-based therapies which involve administering of anti-polypeptides and anti-polynucleotide antibodies to a mammalian, preferably human, patient for treating one or more of the described disorders. Methods for producing anti-polypeptides and anti-polynucleotide antibodies polyclonal and monoclonal antibodies are described in detail elsewhere herein. Such antibodies may be provided in pharmaceutically acceptable compositions as known in the art or as described herein.

A summary of the ways in which the antibodies of the present invention may be used therapeutically includes binding polynucleotides or polypeptides of the present invention locally or systemically in the body or by direct cytotoxicity of the antibody, e.g. as mediated by complement (CDC) or by effector cells (ADCC). Some of these approaches are described in more detail below. Armed with the teachings provided herein, one of ordinary skill in the art will know how to use the antibodies of the present invention for diagnostic, monitoring or therapeutic purposes without undue experimentation.

In particular, the antibodies, fragments and derivatives of the present invention are useful for treating a subject having or developing cell proliferative and/or differentiation disorders as described herein. Such treatment comprises administering a single or multiple doses of the antibody, or a fragment, derivative, or a conjugate thereof.

The antibodies of this invention may be advantageously utilized in combination with other monoclonal or chimeric antibodies, or with lymphokines or hematopoietic growth factors, for example, which serve to increase the number or activity of effector cells which interact with the antibodies.

It is preferred to use high affinity and/or potent in vivo inhibiting and/or neutralizing antibodies against polypeptides or polynucleotides of the present invention, fragments or regions thereof, for both immunoassays directed to and therapy of disorders related to polynucleotides or polypeptides, including fragments thereof, of the present invention. Such antibodies, fragments, or regions, will preferably have an affinity for polynucleotides or polypeptides, including fragments thereof. Preferred binding affinities include those with a dissociation constant or Kd less than 5×10⁻⁶M, 10⁻⁶M, 5×10⁻⁷M, 10⁻⁷M, 5×10⁻⁸M, 10⁻⁸M, 5×10⁻⁹M, 10⁻⁹M, 5×10⁻¹⁰M, 10⁻¹⁰M, 5×10⁻¹¹M, 10⁻¹¹M, 5×10⁻¹²M, 10⁻¹²M, 5×10⁻¹³M, 10⁻¹³M, 5×10⁻¹⁴M, 10⁻¹⁴M, 5×10⁻¹⁵M, and 10⁻¹⁵M.

Moreover, polypeptides of the present invention are useful in inhibiting the angiogenesis of proliferative cells or tissues, either alone, as a protein fusion, or in combination with other polypeptides directly or indirectly, as described elsewhere herein. In a most preferred embodiment, said anti-angiogenesis effect may be achieved indirectly, for example, through the inhibition of hematopoietic, tumor-specific cells, such as tumor-associated macrophages (See Joseph I B, et al. J Natl Cancer Inst, 90(21):1648-53 (1998), which is hereby incorporated by reference). Antibodies directed to polypeptides or polynucleotides of the present invention may also result in inhibition of angiogenesis directly, or indirectly (See Witte L, et al., Cancer Metastasis Rev. 17(2):155-61 (1998), which is hereby incorporated by reference)).

Polypeptides, including protein fusions, of the present invention, or fragments thereof may be useful in inhibiting proliferative cells or tissues through the induction of apoptosis. Said polypeptides may act either directly, or indirectly to induce apoptosis of proliferative cells and tissues, for example in the activation of a death-domain receptor, such as tumor necrosis factor (TNF) receptor-1, CD95 (Fas/APO-1), TNF-receptor-related apoptosis-mediated protein (TRAMP) and TNF-related apoptosis-inducing ligand (TRAIL) receptor-1 and -2 (See Schulze-Osthoff K, et.al., Eur J Biochem 254(3):439-59 (1998), which is hereby incorporated by reference). Moreover, in another preferred embodiment of the present invention, said polypeptides may induce apoptosis through other mechanisms, such as in the activation of other proteins which will activate apoptosis, or through stimulating the expression of said proteins, either alone or in combination with small molecule drugs or adjuvants, such as apoptonin, galectins, thioredoxins, anti-inflammatory proteins (See for example, Mutat Res 400(1-2):447-55 (1998), Mcd Hypotheses. 50(5):423-33 (1998), Chem Biol Interact. April 24; 111-112:23-34 (1998), J Mol Med. 76(6):402-12 (1998), Int J Tissue React; 20(1):3-15 (1998), which are all hereby incorporated by reference).

Polypeptides, including protein fusions to, or fragments thereof, of the present invention are useful in inhibiting the metastasis of proliferative cells or tissues. Inhibition may occur as a direct result of administering polypeptides, or antibodies directed to said polypeptides as described elsewere herein, or indirectly, such as activating the expression of proteins known to inhibit metastasis, for example alpha 4 integrins, (See, e.g., Curr Top Microbiol Immunol 1998; 231:125-41, which is hereby incorporated by reference). Such therapeutic affects of the present invention may be achieved either alone, or in combination with small molecule drugs or adjuvants.

In another embodiment, the invention provides a method of delivering compositions containing the polypeptides of the invention (e.g., compositions containing polypeptides or polypeptide antibodies associated with heterologous polypeptides, heterologous nucleic acids, toxins, or prodrugs) to targeted cells expressing the polypeptide of the present invention. Polypeptides or polypeptide antibodies of the invention may be associated with with heterologous polypeptides, heterologous nucleic acids, toxins, or prodrugs via hydrophobic, hydrophilic, ionic and/or covalent interactions.

Polypeptides, protein fusions to, or fragments thereof, of the present invention are useful in enhancing the immunogenicity and/or antigenicity of proliferating cells or tissues, either directly, such as would occur if the polypeptides of the present invention ‘vaccinated’ the immune response to respond to proliferative antigens and immunogens, or indirectly, such as in activating the expression of proteins known to enhance the immune response (e.g. chemokines), to said antigens and immunogens.

Renal Disorders

Polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention, may be used to treat, prevent, diagnose, and/or prognose disorders of the renal system. Renal disorders which can be diagnosed, prognosed, prevented, and/or treated with compositions of the invention include, but are not limited to, kidney failure, nephritis, blood vessel disorders of kidney, metabolic and congenital kidney disorders, urinary disorders of the kidney, autoimmune disorders, sclerosis and necrosis, electrolyte imbalance, and kidney cancers.

Kidney diseases which can be diagnosed, prognosed, prevented, and/or treated with compositions of the invention include, but are not limited to, acute kidney failure, chronic kidney failure, atheroembolic renal failure, end-stage renal disease, inflammatory diseases of the kidney (e.g., acute glomerulonephritis, postinfectious glomerulonephritis, rapidly progressive glomerulonephritis, nephrotic syndrome, membranous glomerulonephritis, familial nephrotic syndrome, membranoproliferative glomerulonephritis I and II, mesangial proliferative glomerulonephritis, chronic glomerulonephritis, acute tubulointerstitial nephritis, chronic tubulointerstitial nephritis, acute post-streptococcal glomerulonephritis (PSGN), pyelonephritis, lupus nephritis, chronic nephritis, interstitial nephritis, and post-streptococcal glomerulonephritis), blood vessel disorders of the kidneys (e.g., kidney infarction, atheroembolic kidney disease, cortical necrosis, malignant nephrosclerosis, renal vein thrombosis, renal underperfusion, renal retinopathy, renal ischemia-reperfusion, renal artery embolism, and renal artery stenosis), and kidney disorders resulting form urinary tract disease (e.g., pyelonephritis, hydronephrosis, urolithiasis (renal lithiasis, nephrolithiasis), reflux nephropathy, urinary tract infections, urinary retention, and acute or chronic unilateral obstructive uropathy.)

In addition, compositions of the invention can be used to diagnose, prognose, prevent, and/or treat metabolic and congenital disorders of the kidney (e.g., uremia, renal amyloidosis, renal osteodystrophy, renal tubular acidosis, renal glycosuria, nephrogenic diabetes insipidus, cystinuria, Fanconi's syndrome, renal fibrocystic osteosis (renal rickets), Haitnup disease, Bartter's syndrome, Liddle's syndrome, polycystic kidney disease, medullary cystic disease, medullary sponge kidney, Alport's syndrome, nail-patella syndrome, congenital nephrotic syndrome, CRUSH syndrome, horseshoe kidney, diabetic nephropathy, nephrogenic diabetes insipidus, analgesic nephropathy, kidney stones, and membranous nephropathy), and autoimmune disorders of the kidney (e.g., systemic lupus erythematosus (SLE), Goodpasture syndrome, IgA nephropathy, and IgM mesangial proliferative glomerulonephritis).

Compositions of the invention can also be used to diagnose, prognose, prevent, and/or treat sclerotic or necrotic disorders of the kidney (e.g., glomerulosclerosis, diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), necrotizing glomerulonephritis, and renal papillary necrosis), cancers of the kidney (e.g., nephroma, hypernephroma, nephroblastoma, renal cell cancer, transitional cell cancer, renal adenocarcinoma, squamous cell cancer, and Wilm's tumor), and electrolyte imbalances (e.g., nephrocalcinosis, pyuria, edema, hydronephritis, proteinuria, hyponatremia, hypernatremia, hypokalemia, hyperkalemia, hypocalcemia, hypercalcemia, hypophosphatemia, and hyperphosphatemia).

Polypeptides may be administered using any method known in the art, including, but not limited to, direct needle injection at the delivery site, intravenous injection, topical administration, catheter infusion, biolistic injectors, particle accelerators, gelfoam sponge depots, other commercially available depot materials, osmotic pumps, oral or suppositorial solid pharmaceutical formulations, decanting or topical applications during surgery, aerosol delivery. Such methods are known in the art. Polypeptides may be administered as part of a Therapeutic, described in more detail below. Methods of delivering polynucleotides are described in more detail herein.

Cardiovascular Disorders

Polynucleotides or polypeptides, or agonists or antagonists of the present invention, may be used to treat, prevent, diagnose, and/or prognose cardiovascular disorders, including, but not limited to, peripheral artery disease, such as limb ischemia.

Cardiovascular disorders include, but are not limited to, cardiovascular abnormalities, such as arterio-arterial fistula, arteriovenous fistula, cerebral arteriovenous malformations, congenital heart defects, pulmonary atresia, and Scimitar Syndrome. Congenital heart defects include, but are not limited to, aortic coarctation, cor triatriatum, coronary vessel anomalies, crisscross heart, dextrocardia, patent ductus arteriosus, Ebstein's anomaly, Eisenmenger complex, hypoplastic left heart syndrome, levocardia, tetralogy of fallot, transposition of great vessels, double outlet right ventricle, tricuspid atresia, persistent truncus arteriosus, and heart septal defects, such as aortopulmonary septal defect, endocardial cushion defects, Lutembacher's Syndrome, trilogy of Fallot, ventricular heart septal defects.

Cardiovascular disorders also include, but are not limited to, heart disease, such as arrhythmias, carcinoid heart disease, high cardiac output, low cardiac output, cardiac tamponade, endocarditis (including bacterial), heart aneurysm, cardiac arrest, congestive heart failure, congestive cardiomyopathy, paroxysmal dyspnea, cardiac edema, heart hypertrophy, congestive cardiomyopathy, left ventricular hypertrophy, right ventricular hypertrophy, post-infarction heart rupture, ventricular septal rupture, heart valve diseases, myocardial diseases, myocardial ischemia, pericardial effusion, pericarditis (including constrictive and tuberculous), pneumopericardium, postpericardiotomy syndrome, pulmonary heart disease, rheumatic heart disease, ventricular dysfunction, hyperemia, cardiovascular pregnancy complications, Scimitar Syndrome, cardiovascular syphilis, and cardiovascular tuberculosis.

Arrhythmias include, but are not limited to, sinus arrhythmia, atrial fibrillation, atrial flutter, bradycardia, extrasystole, Adams-Stokes Syndrome, bundle-branch block, sinoatrial block, long QT syndrome, parasystole, Lown-Ganong-Levine Syndrome, Mahaim-type pre-excitation syndrome, Wolff-Parkinson-White syndrome, sick sinus syndrome, tachycardias, and ventricular fibrillation. Tachycardias include paroxysmal tachycardia, supraventricular tachycardia, accelerated idioventricular rhythm, atrioventricular nodal reentry tachycardia, ectopic atrial tachycardia, ectopic junctional tachycardia, sinoatrial nodal reentry tachycardia, sinus tachycardia, Torsades de Pointes, and ventricular tachycardia.

Heart valve diseases include, but are not limited to, aortic valve insufficiency, aortic valve stenosis, hear murmurs, aortic valve prolapse, mitral valve prolapse, tricuspid valve prolapse, mitral valve insufficiency, mitral valve stenosis, pulmonary atresia, pulmonary valve insufficiency, pulmonary valve stenosis, tricuspid atresia, tricuspid valve insufficiency, and tricuspid valve stenosis.

Myocardial diseases include, but are not limited to, alcoholic cardiomyopathy, congestive cardiomyopathy, hypertrophic cardiomyopathy, aortic subvalvular stenosis, pulmonary subvalvular stenosis, restrictive cardiomyopathy, Chagas cardiomyopathy, endocardial fibroelastosis, endomyocardial fibrosis, Kearns Syndrome, myocardial reperfusion injury, and myocarditis.

Myocardial ischemias include, but are not limited to, coronary disease, such as angina pectoris, coronary aneurysm, coronary arteriosclerosis, coronary thrombosis, coronary vasospasm, myocardial infarction and myocardial stunning.

Cardiovascular diseases also include vascular diseases such as aneurysms, angiodysplasia, angiomatosis, bacillary angiomatosis, Hippel-Lindau Disease, Klippel-Trenaunay-Weber Syndrome, Sturge-Weber Syndrome, angioneurotic edema, aortic diseases, Takayasu's Arteritis, aortitis, Leriche's Syndrome, arterial occlusive diseases, arteritis, enarteritis, polyarteritis nodosa, cerebrovascular disorders, diabetic angiopathies, diabetic retinopathy, embolisms, thrombosis, erythromelalgia, hemorrhoids, hepatic veno-occlusive disease, hypertension, hypotension, ischemia, peripheral vascular diseases, phlebitis, pulmonary veno-occlusive disease, Raynaud's disease, CREST syndrome, retinal vein occlusion, Scimitar syndrome, superior vena cava syndrome, telangiectasia, atacia telangiectasia, hereditary hemorrhagic telangiectasia, varicocele, varicose veins, varicose ulcer, vasculitis, and venous insufficiency.

Aneurysms include, but are not limited to, dissecting aneurysms, false aneurysms, infected aneurysms, ruptured aneurysms, aortic aneurysms, cerebral aneurysms, coronary aneurysms, heart aneurysms, and iliac aneurysms.

Arterial occlusive diseases include, but are not limited to, arteriosclerosis, intermittent claudication, carotid stenosis, fibromuscular dysplasias, mesenteric vascular occlusion, Moyamoya disease, renal artery obstruction, retinal artery occlusion, and thromboangiitis obliterans.

Cerebrovascular disorders include, but are not limited to, carotid artery diseases, cerebral amyloid angiopathy, cerebral aneurysm, cerebral anoxia, cerebral arteriosclerosis, cerebral arteriovenous malformation, cerebral artery diseases, cerebral embolism and thrombosis, carotid artery thrombosis, sinus thrombosis, Wallenberg's syndrome, cerebral hemorrhage, epidural hematoma, subdural hematoma, subaraxhnoid hemorrhage, cerebral infarction, cerebral ischemia (including transient), subclavian steal syndrome, periventricular leukomalacia, vascular headache, cluster headache, migraine, and vertebrobasilar insufficiency.

Embolisms include, but are not limited to, air embolisms, amniotic fluid embolisms, cholesterol embolisms, blue toe syndrome, fat embolisms, pulmonary embolisms, and thromoboembolisms. Thrombosis include, but are not limited to, coronary thrombosis, hepatic vein thrombosis, retinal vein occlusion, carotid artery thrombosis, sinus thrombosis, Wallenberg's syndrome, and thrombophlebitis.

Ischemic disorders include, but are not limited to, cerebral ischemia, ischemic colitis, compartment syndromes, anterior compartment syndrome, myocardial ischemia, reperfusion injuries, and peripheral limb ischemia. Vasculitis includes, but is not limited to, aortitis, arteritis, Behcet's Syndrome, Churg-Strauss Syndrome, mucocutaneous lymph node syndrome, thromboangiitis obliterans, hypersensitivity vasculitis, Schoenlein-Henoch purpura, allergic cutaneous vasculitis, and Wegener's granulomatosis.

Polypeptides may be administered using any method known in the art, including, but not limited to, direct needle injection at the delivery site, intravenous injection, topical administration, catheter infusion, biolistic injectors, particle accelerators, gelfoam sponge depots, other commercially available depot materials, osmotic pumps, oral or suppositorial solid pharmaceutical formulations, decanting or topical applications during surgery, aerosol delivery. Such methods are known in the art. Polypeptides may be administered as part of a Therapeutic, described in more detail below. Methods of delivering polynucleotides are described in more detail herein.

Respiratory Disorders

Polynucleotides or polypeptides, or agonists or antagonists of the present invention may be used to treat, prevent, diagnose, and/or prognose diseases and/or disorders of the respiratory system.

Diseases and disorders of the respiratory system include, but are not limited to, nasal vestibulitis, nonallergic rhinitis (e.g., acute rhinitis, chronic rhinitis, atrophic rhinitis, vasomotor rhinitis), nasal polyps, and sinusitis, juvenile angiofibromas, cancer of the nose and juvenile papillomas, vocal cord polyps, nodules (singer's nodules), contact ulcers, vocal cord paralysis, laryngoceles, pharyngitis (e.g., viral and bacterial), tonsillitis, tonsillar cellulitis, parapharyngeal abscess, laryngitis, laryngoceles, and throat cancers (e.g., cancer of the nasopharynx, tonsil cancer, larynx cancer), lung cancer (e.g., squamous cell carcinoma, small cell (oat cell) carcinoma, large cell carcinoma, and adenocarcinoma), allergic disorders (eosinophilic pneumonia, hypersensitivity pneumonitis (e.g., extrinsic allergic alveolitis, allergic interstitial pneumonitis, organic dust pneumoconiosis, allergic bronchopulmonary aspergillosis, asthma, Wegener's granulomatosis (granulomatous vasculitis), Goodpasture's syndrome)), pneumonia (e.g., bacterial pneumonia (e.g., Streptococcus pneumoniae (pneumoncoccal pneumonia), Staphylococcus aureus (staphylococcal pneumonia), Gram-negative bacterial pneumonia (caused by, e.g., Klebsiella and Pseudomas spp.), Mycoplasma pneumoniae pneumonia, Hemophilus influenzae pneumonia, Legionella pneumophila (Legionnaires' disease), and Chlamydia psittaci (Psittacosis)), and viral pneumonia (e.g., influenza, chickenpox (varicella).

Additional diseases and disorders of the respiratory system include, but are not limited to bronchiolitis, polio (poliomyelitis), croup, respiratory syncytial viral infection, mumps, erythema infectiosum (fifth disease), roseola infantum, progressive rubella panencephalitis, german measles, and subacute sclerosing panencephalitis), fungal pneumonia (e.g., Histoplasmosis, Coccidioidomycosis, Blastomycosis, fungal infections in people with severely suppressed immune systems (e.g., cryptococcosis, caused by Cryptococcus neoformans; aspergillosis, caused by Aspergillus spp.; candidiasis, caused by Candida; and mucormycosis)), Pneumocystis carinii (pneumocystis pneumonia), atypical pneumonias (e.g., Mycoplasma and Chlamydia spp.), opportunistic infection pneumonia, nosocomial pneumonia, chemical pneumonitis, and aspiration pneumonia, pleural disorders (e.g., pleurisy, pleural effusion, and pneumothorax (e.g., simple spontaneous pneumothorax, complicated spontaneous pneumothorax, tension pneumothorax)), obstructive airway diseases (e.g., asthma, chronic obstructive pulmonary disease (COPD), emphysema, chronic or acute bronchitis), occupational lung diseases (e.g., silicosis, black lung (coal workers' pneumoconiosis), asbestosis, berylliosis, occupational asthsma, byssinosis, and benign pneumoconioses), Infiltrative Lung Disease (e.g., pulmonary fibrosis (e.g., fibrosing alveolitis, usual interstitial pneumonia), idiopathic pulmonary fibrosis, desquamative interstitial pneumonia, lymphoid interstitial pneumonia, histiocytosis X (e.g., Letterer-Siwe disease, Hand-Schüller-Christian disease, cosinophilic granuloma), idiopathic pulmonary hemosiderosis, sarcoidosis and pulmonary alveolar proteinosis), Acute respiratory distress syndrome (also called, e.g., adult respiratory distress syndrome), edema, pulmonary embolism, bronchitis (e.g., viral, bacterial), bronchiectasis, atelectasis, lung abscess (caused by, e.g., Staphylococcus aureus or Legionella pneumophila), and cystic fibrosis.

Anti-Angiogenesis Activity

The naturally occurring balance between endogenous stimulators and inhibitors of angiogenesis is one in which inhibitory influences predominate. Rastinejad et al., Cell 56:345-355 (1989). In those rare instances in which neovascularization occurs under normal physiological conditions, such as wound healing, organ regeneration, embryonic development, and female reproductive processes, angiogenesis is stringently regulated and spatially and temporally delimited. Under conditions of pathological angiogenesis such as that characterizing solid tumor growth, these regulatory controls fail. Unregulated angiogenesis becomes pathologic and sustains progression of many neoplastic and non-neoplastic diseases. A number of serious diseases are dominated by abnormal neovascularization including solid tumor growth and metastases, arthritis, some types of eye disorders, and psoriasis. See, e.g., reviews by Moses et al., Biotech. 9:630-634 (1991); Folkman et al., N. Engl. J. Med., 333:1757-1763 (1995); Auerbach et al., J. Microvasc. Res. 29:401-411 (1985); Folkman, Advances in Cancer Research, eds. Klein and Weinhouse, Academic Press, New York, pp. 175-203 (1985); Patz, Am. J. Opthalmol. 94:715-743 (1982); and Folkman et al., Science 221:719-725 (1983). In a number of pathological conditions, the process of angiogenesis contributes to the disease state. For example, significant data have accumulated which suggest that the growth of solid tumors is dependent on angiogenesis. Folkman and Klagsbrun, Science 235:442-447 (1987).

The present invention provides for treatment of diseases or disorders associated with neovascularization by administration of the polynucleotides and/or polypeptides of the invention, as well as agonists or antagonists of the present invention. Malignant and metastatic conditions which can be treated with the polynucleotides and polypeptides, or agonists or antagonists of the invention include, but are not limited to, malignancies, solid tumors, and cancers described herein and otherwise known in the art (for a review of such disorders, see Fishman et al., Medicine, 2d Ed., J. B. Lippincott Co., Philadelphia (1985)). Thus, the present invention provides a method of treating an angiogenesis-related disease and/or disorder, comprising administering to an individual in need thereof a therapeutically effective amount of a polynucleotide, polypeptide, antagonist and/or agonist of the invention. For example, polynucleotides, polypeptides, antagonists and/or agonists may be utilized in a variety of additional methods in order to therapeutically treat a cancer or tumor. Cancers which may be treated with polynucleotides, polypeptides, antagonists and/or agonists include, but are not limited to solid tumors, including prostate, lung, breast, ovarian, stomach, pancreas, larynx, esophagus, testes, liver, parotid, biliary tract, colon, rectum, cervix, uterus, endometrium, kidney, bladder, thyroid cancer; primary tumors and metastases; melanomas; glioblastoma; Kaposi's sarcoma; leiomyosarcoma; non-small cell lung cancer; colorectal cancer; advanced malignancies; and blood born tumors such as leukemias. For example, polynucleotides, polypeptides, antagonists and/or agonists may be delivered topically, in order to treat cancers such as skin cancer, head and neck tumors, breast tumors, and Kaposi's sarcoma.

Within yet other aspects, polynucleotides, polypeptides, antagonists and/or agonists may be utilized to treat superficial forms of bladder cancer by, for example, intravesical administration. Polynucleotides, polypeptides, antagonists and/or agonists may be delivered directly into the tumor, or near the tumor site, via injection or a catheter. Of course, as the artisan of ordinary skill will appreciate, the appropriate mode of administration will vary according to the cancer to be treated. Other modes of delivery are discussed herein.

Polynucleotides, polypeptides, antagonists and/or agonists may be useful in treating other disorders, besides cancers, which involve angiogenesis. These disorders include, but are not limited to: benign tumors, for example hemangiomas, acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas; artheroscleric plaques; ocular angiogenic diseases, for example, diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis, retinoblastoma, uvietis and Pterygia (abnormal blood vessel growth) of the eye; rheumatoid arthritis; psoriasis; delayed wound healing; endometriosis; vasculogenesis; granulations; hypertrophic scars (keloids); nonunion fractures; scleroderma; trachoma; vascular adhesions; myocardial angiogenesis; coronary collaterals; cerebral collaterals; arteriovenous malformations; ischemic limb angiogenesis; Osler-Webber Syndrome; plaque neovascularization; telangiectasia; hemophiliac joints; angiofibroma; fibromuscular dysplasia; wound granulation; Crohn's disease; and atherosclerosis.

For example, within one aspect of the present invention methods are provided for treating hypertrophic scars and keloids, comprising the step of administering a polynucleotide, polypeptide, antagonist and/or agonist of the invention to a hypertrophic scar or keloid.

Within one embodiment of the present invention polynucleotides, polypeptides, antagonists and/or agonists of the invention are directly injected into a hypertrophic scar or keloid, in order to prevent the progression of these lesions. This therapy is of particular value in the prophylactic treatment of conditions which are known to result in the development of hypertrophic scars and keloids (e.g., burns), and is preferably initiated after the proliferative phase has had time to progress (approximately 14 days after the initial injury), but before hypertrophic scar or keloid development. As noted above, the present invention also provides methods for treating neovascular diseases of the eye, including for example, corneal neovascularization, neovascular glaucoma, proliferative diabetic retinopathy, retrolental fibroplasia and macular degeneration.

Moreover, Ocular disorders associated with neovascularization which can be treated with the polynucleotides and polypeptides of the present invention (including agonists and/or antagonists) include, but are not limited to: neovascular glaucoma, diabetic retinopathy, retinoblastoma, retrolental fibroplasia, uveitis, retinopathy of prematurity macular degeneration, corneal graft neovascularization, as well as other eye inflammatory diseases, ocular tumors and diseases associated with choroidal or iris neovascularization. See, e.g., reviews by Waltman et al., Am. J. Ophthal. 85:704-710 (1978) and Gartner et al., Surv. Ophthal. 22:291-312 (1978).

Thus, within one aspect of the present invention methods are provided for treating neovascular diseases of the eye such as corneal neovascularization (including corneal graft neovascularization), comprising the step of administering to a patient a therapeutically effective amount of a compound (as described above) to the cornea, such that the formation of blood vessels is inhibited. Briefly, the cornea is a tissue which normally lacks blood vessels. In certain pathological conditions however, capillaries may extend into the cornea from the pericorneal vascular plexus of the limbus. When the cornea becomes vascularized, it also becomes clouded, resulting in a decline in the patient's visual acuity. Visual loss may become complete if the cornea completely opacitates. A wide variety of disorders can result in corneal neovascularization, including for example, corneal infections (e.g., trachoma, herpes simplex keratitis, leishmaniasis and onchocerciasis), immunological processes (e.g., graft rejection and Stevens-Johnson's syndrome), alkali burns, trauma, inflammation (of any cause), toxic and nutritional deficiency states, and as a complication of wearing contact lenses.

Within particularly preferred embodiments of the invention, may be prepared for topical administration in saline (combined with any of the preservatives and antimicrobial agents commonly used in ocular preparations), and administered in eyedrop form. The solution or suspension may be prepared in its pure form and administered several times daily. Alternatively, anti-angiogenic compositions, prepared as described above, may also be administered directly to the cornea. Within preferred embodiments, the anti-angiogenic composition is prepared with a muco-adhesive polymer which binds to cornea. Within further embodiments, the anti-angiogenic factors or anti-angiogenic compositions may be utilized as an adjunct to conventional steroid therapy. Topical therapy may also be useful prophylactically in corneal lesions which are known to have a high probability of inducing an angiogenic response (such as chemical burns). In these instances the treatment, likely in combination with steroids, may be instituted immediately to help prevent subsequent complications.

Within other embodiments, the compounds described above may be injected directly into the corneal stroma by an ophthalmologist under microscopic guidance. The preferred site of injection may vary with the morphology of the individual lesion, but the goal of the administration would be to place the composition at the advancing front of the vasculature (i.e., interspersed between the blood vessels and the normal cornea). In most cases this would involve perilimbic corneal injection to “protect” the cornea from the advancing blood vessels. This method may also be utilized shortly after a corneal insult in order to prophylactically prevent corneal neovascularization. In this situation the material could be injected in the perilimbic cornea interspersed between the corneal lesion and its undesired potential limbic blood supply. Such methods may also be utilized in a similar fashion to prevent capillary invasion of transplanted corneas. In a sustained-release form injections might only be required 2-3 times per year. A steroid could also be added to the injection solution to reduce inflammation resulting from the injection itself.

Within another aspect of the present invention, methods are provided for treating neovascular glaucoma, comprising the step of administering to a patient a therapeutically effective amount of a polynucleotide, polypeptide, antagonist and/or agonist to the eye, such that the formation of blood vessels is inhibited. In one embodiment, the compound may be administered topically to the eye in order to treat early forms of neovascular glaucoma. Within other embodiments, the compound may be implanted by injection into the region of the anterior chamber angle. Within other embodiments, the compound may also be placed in any location such that the compound is continuously released into the aqueous humor. Within another aspect of the present invention, methods are provided for treating proliferative diabetic retinopathy, comprising the step of administering to a patient a therapeutically effective amount of a polynucleotide, polypeptide, antagonist and/or agonist to the eyes, such that the formation of blood vessels is inhibited.

Within particularly preferred embodiments of the invention, proliferative diabetic retinopathy may be treated by injection into the aqueous humor or the vitreous, in order to increase the local concentration of the polynucleotide, polypeptide, antagonist and/or agonist in the retina. Preferably, this treatment should be initiated prior to the acquisition of severe disease requiring photocoagulation.

Within another aspect of the present invention, methods are provided for treating retrolental fibroplasia, comprising the step of administering to a patient a therapeutically effective amount of a polynucleotide, polypeptide, antagonist and/or agonist to the eye, such that the formation of blood vessels is inhibited. The compound may be administered topically, via intravitreous injection and/or via intraocular implants.

Additionally, disorders which can be treated with the polynucleotides, polypeptides, agonists and/or agonists include, but are not limited to, hemangioma, arthritis, psoriasis, angiofibroma, atherosclerotic plaques, delayed wound healing, granulations, hemophilic joints, hypertrophic scars, nonunion fractures, Osler-Weber syndrome, pyogenic granuloma, scleroderma, trachoma, and vascular adhesions.

Moreover, disorders and/or states, which can be treated, prevented, diagnosed, and/or prognosed with the polynucleotides, polypeptides, agonists and/or agonists of the invention include, but are not limited to, solid tumors, blood born tumors such as leukemias, tumor metastasis, Kaposi's sarcoma, benign tumors, for example hemangiomas, acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas, rheumatoid arthritis, psoriasis, ocular angiogenic diseases, for example, diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis, retinoblastoma, and uvietis, delayed wound healing, endometriosis, vascluogenesis, granulations, hypertrophic scars (keloids), nonunion fractures, scleroderma, trachoma, vascular adhesions, myocardial angiogenesis, coronary collaterals, cerebral collaterals, arteriovenous malformations, ischemic limb angiogenesis, Osler-Webber Syndrome, plaque neovascularization, telangiectasia, hemophiliac joints, angiofibroma fibromuscular dysplasia, wound granulation, Crohn's disease, atherosclerosis, birth control agent by preventing vascularization required for embryo implantation controlling menstruation, diseases that have angiogenesis as a pathologic consequence such as cat scratch disease (Rochele minalia quintosa), ulcers (Helicobacter pylori), Bartonellosis and bacillary angiomatosis.

In one aspect of the birth control method, an amount of the compound sufficient to block embryo implantation is administered before or after intercourse and fertilization have occurred, thus providing an effective method of birth control, possibly a “morning after” method. Polynucleotides, polypeptides, agonists and/or agonists may also be used in controlling menstruation or administered as either a peritoneal lavage fluid or for peritoneal implantation in the treatment of endometriosis.

Polynucleotides, polypeptides, agonists and/or agonists of the present invention may be incorporated into surgical sutures in order to prevent stitch granulomas.

Polynucleotides, polypeptides, agonists and/or agonists may be utilized in a wide variety of surgical procedures. For example, within one aspect of the present invention a compositions (in the form of, for example, a spray or film) may be utilized to coat or spray an area prior to removal of a tumor, in order to isolate normal surrounding tissues from malignant tissue, and/or to prevent the spread of disease to surrounding tissues. Within other aspects of the present invention, compositions (e.g., in the form of a spray) may be delivered via endoscopic procedures in order to coat tumors, or inhibit angiogenesis in a desired locale. Within yet other aspects of the present invention, surgical meshes which have been coated with anti-angiogenic compositions of the present invention may be utilized in any procedure wherein a surgical mesh might be utilized. For example, within one embodiment of the invention a surgical mesh laden with an anti-angiogenic composition may be utilized during abdominal cancer resection surgery (e.g., subsequent to colon resection) in order to provide support to the structure, and to release an amount of the anti-angiogenic factor.

Within further aspects of the present invention, methods are provided for treating tumor excision sites, comprising administering a polynucleotide, polypeptide, agonist and/or agonist to the resection margins of a tumor subsequent to excision, such that the local recurrence of cancer and the formation of new blood vessels at the site is inhibited. Within one embodiment of the invention, the anti-angiogenic compound is administered directly to the tumor excision site (e.g., applied by swabbing, brushing or otherwise coating the resection margins of the tumor with the anti-angiogenic compound). Alternatively, the anti-angiogenic compounds may be incorporated into known surgical pastes prior to administration. Within particularly preferred embodiments of the invention, the anti-angiogenic compounds are applied after hepatic resections for malignancy, and after neurosurgical operations.

Within one aspect of the present invention, polynucleotides, polypeptides, agonists and/or agonists may be administered to the resection margin of a wide variety of tumors, including for example, breast, colon, brain and hepatic tumors. For example, within one embodiment of the invention, anti-angiogenic compounds may be administered to the site of a neurological tumor subsequent to excision, such that the formation of new blood vessels at the site are inhibited.

The polynucleotides, polypeptides, agonists and/or agonists of the present invention may also be administered along with other anti-angiogenic factors. Representative examples of other anti-angiogenic factors include: Anti-Invasive Factor, retinoic acid and derivatives thereof, paclitaxel, Suramin, Tissue Inhibitor of Metalloproteinase-1, Tissue Inhibitor of Metalloproteinase-2, Plasminogen Activator Inhibitor-1, Plasminogen Activator Inhibitor-2, and various forms of the lighter “d group” transition metals.

Lighter “d group” transition metals include, for example, vanadium, molybdenum, tungsten, titanium, niobium, and tantalum species. Such transition metal species may form transition metal complexes. Suitable complexes of the above-mentioned transition metal species include oxo transition metal complexes.

Representative examples of vanadium complexes include oxo vanadium complexes such as vanadate and vanadyl complexes. Suitable vanadate complexes include metavanadate and orthovanadate complexes such as, for example, ammonium metavanadate, sodium metavanadate, and sodium orthovanadate. Suitable vanadyl complexes include, for example, vanadyl acetylacetonate and vanadyl sulfate including vanadyl sulfate hydrates such as vanadyl sulfate mono- and trihydrates.

Representative examples of tungsten and molybdenum complexes also include oxo complexes. Suitable oxo tungsten complexes include tungstate and tungsten oxide complexes. Suitable tungstate complexes include ammonium tungstate, calcium tungstate, sodium tungstate dihydrate, and tungstic acid. Suitable tungsten oxides include tungsten (IV) oxide and tungsten (VI) oxide. Suitable oxo molybdenum complexes include molybdate, molybdenum oxide, and molybdenyl complexes. Suitable molybdate complexes include ammonium molybdate and its hydrates, sodium molybdate and its hydrates, and potassium molybdate and its hydrates. Suitable molybdenum oxides include molybdenum (VI) oxide, molybdenum (VI) oxide, and molybdic acid. Suitable molybdenyl complexes include, for example, molybdenyl acetylacetonate. Other suitable tungsten and molybdenum complexes include hydroxo derivatives derived from, for example, glycerol, tartaric acid, and sugars.

A wide variety of other anti-angiogenic factors may also be utilized within the context of the present invention. Representative examples include platelet factor 4; protamine sulphate; sulphated chitin derivatives (prepared from queen crab shells), (Murata et al., Cancer Res. 51:22-26, 1991); Sulphated Polysaccharide Peptidoglycan Complex (SP-PG) (the function of this compound may be enhanced by the presence of steroids such as estrogen, and tamoxifen citrate); Staurosporine; modulators of matrix metabolism, including for example, proline analogs, cishydroxyproline, d,L-3,4-dehydroproline, Thiaproline, alpha,alpha-dipyridyl, aminopropionitrile fumarate; 4-propyl-5-(4-pyridinyl)-2(3H)-oxazolone; Methotrexate; Mitoxantrone; Heparin; Interferons; 2 Macroglobulin-serum; ChIMP-3 (Pavioff et al., J. Bio. Chem. 267:17321-17326, 1992); Chymostatin (Tomkinson et al., Biochem J. 286:475-480, 1992); Cyclodextrin Tetradecasulfate; Eponemycin; Camptothecin; Fumagillin (Ingber et al., Nature 348:555-557, 1990); Gold Sodium Thiomalate (“GST”; Matsubara and Ziff, J. Clin. Invest. 79:1440-1446, 1987); anticollagenase-serum; alpha2-antiplasmin (Holmes et al., J. Biol. Chem. 262(4):1659-1664, 1987); Bisantrene (National Cancer Institute); Lobenzarit disodium (N-(2)-carboxyphenyl-4-chloroanthronilic acid disodium or “CCA”; Takeuchi et al., Agents Actions 36:312-316, 1992); Thalidomide; Angostatic steroid; AGM-1470; carboxynaminolmidazole; and metalloproteinase inhibitors such as BB94.

Diseases at the Cellular Level

Diseases associated with increased cell survival or the inhibition of apoptosis that could be treated, prevented, diagnosed, and/or prognosed using polynucleotides or polypeptides, as well as antagonists or agonists of the present invention, include cancers (such as follicular lymphomas, carcinomas with p53 mutations, and hormone-dependent tumors, including, but not limited to colon cancer, cardiac tumors, pancreatic cancer, melanoma, retinoblastoma, glioblastoma, lung cancer, intestinal cancer, testicular cancer, stomach cancer, neuroblastoma, myxoma, myoma, lymphoma, endothelioma, osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, adenoma, breast cancer, prostate cancer, Kaposi's sarcoma and ovarian cancer); autoimmune disorders (such as, multiple sclerosis, Sjogren's syndrome, Hashimoto's thyroiditis, biliary cirrhosis, Behcet's disease, Crohn's disease, polymyositis, systemic lupus erythematosus and immune-related glomerulonephritis and rheumatoid arthritis) and viral infections (such as herpes viruses, pox viruses and adenoviruses), inflammation, graft v. host disease, acute graft rejection, and chronic graft rejection.

In preferred embodiments, polynucleotides, polypeptides, and/or antagonists of the invention are used to inhibit growth, progression, and/or metasis of cancers, in particular those listed above.

Additional diseases or conditions associated with increased cell survival that could be treated or detected by polynucleotides or polypeptides, or agonists or antagonists of the present invention include, but are not limited to, progression, and/or metastases of malignancies and related disorders such as leukemia (including acute leukemias (e.g., acute lymphocytic leukemia, acute myelocytic leukemia (including myeloblastic, promyelocytic, myelomonocytic, monocytic, and erythroleukemia)) and chronic leukemias (e.g., chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia)), polycythemia vera, lymphomas (e.g., Hodgkin's disease and non-Hodgkin's disease), multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, and solid tumors including, but not limited to, sarcomas and carcinomas such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma, and retinoblastoma.

Diseases associated with increased apoptosis that could be treated, prevented, diagnosed, and/or prognosed using polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, include, but are not limited to, AIDS; neurodegenerative disorders (such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Retinitis pigmentosa, Cerebellar degeneration and brain tumor or prior associated disease); autoimmune disorders (such as, multiple sclerosis, Sjogren's syndrome, Hashimoto's thyroiditis, biliary cirrhosis, Behcet's disease, Crohn's disease, polymyositis, systemic lupus erythematosus and immune-related glomerulonephritis and rheumatoid arthritis) myelodysplastic syndromes (such as aplastic anemia), graft v. host disease, ischemic injury (such as that caused by myocardial infarction, stroke and reperfusion injury), liver injury (e.g., hepatitis related liver injury, ischemia/reperfusion injury, cholestosis (bile duct injury) and liver cancer); toxin-induced liver disease (such as that caused by alcohol), septic shock, cachexia and anorexia.

Wound Healing and Epithelial Cell Proliferation

In accordance with yet a further aspect of the present invention, there is provided a process for utilizing polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, for therapeutic purposes, for example, to stimulate epithelial cell proliferation and basal keratinocytes for the purpose of wound healing, and to stimulate hair follicle production and healing of dermal wounds. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, may be clinically useful in stimulating wound healing including surgical wounds, excisional wounds, deep wounds involving damage of the dermis and epidermis, eye tissue wounds, dental tissue wounds, oral cavity wounds, diabetic ulcers, dermal ulcers, cubitus ulcers, arterial ulcers, venous stasis ulcers, burns resulting from heat exposure or chemicals, and other abnormal wound healing conditions such as uremia, malnutrition, vitamin deficiencies and complications associated with systemic treatment with steroids, radiation therapy and antineoplastic drugs and antimetabolites. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to promote dermal reestablishment subsequent to dermal loss

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to increase the adherence of skin grafts to a wound bed and to stimulate re-epithelialization from the wound bed. The following are types of grafts that polynucleotides or polypeptides, agonists or antagonists of the present invention, could be used to increase adherence to a wound bed: autografts, artificial skin, allografts, autodermic graft, autoepdermic grafts, avacular grafts, Blair-Brown grafts, bone graft, brephoplastic grafts, cutis graft, delayed graft, dermic graft, epidermic graft, fascia graft, full thickness graft, heterologous graft, xenograft, homologous graft, hyperplastic graft, lamellar graft, mesh graft, mucosal graft, Ollier-Thiersch graft, omenpal graft, patch graft, pedicle graft, penetrating graft, split skin graft, thick split graft. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, can be used to promote skin strength and to improve the appearance of aged skin.

It is believed that polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, will also produce changes in hepatocyte proliferation, and epithelial cell proliferation in the lung, breast, pancreas, stomach, small intestine, and large intestine. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could promote proliferation of epithelial cells such as sebocytes, hair follicles, hepatocytes, type II pneumocytes, mucin-producing goblet cells, and other epithelial cells and their progenitors contained within the skin, lung, liver, and gastrointestinal tract. Polynucleotides or polypeptides, agonists or antagonists of the present invention, may promote proliferation of endothelial cells, keratinocytes, and basal keratinocytes.

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could also be used to reduce the side effects of gut toxicity that result from radiation, chemotherapy treatments or viral infections. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, may have a cytoprotective effect on the small intestine mucosa. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, may also stimulate healing of mucositis (mouth ulcers) that result from chemotherapy and viral infections.

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could further be used in full regeneration of skin in full and partial thickness skin defects, including burns, (i.e., repopulation of hair follicles, sweat glands, and sebaceous glands), treatment of other skin defects such as psoriasis. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to treat epidermolysis bullosa, a defect in adherence of the epidermis to the underlying dermis which results in frequent, open and painful blisters by accelerating reepithelialization of these lesions. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could also be used to treat gastric and doudenal ulcers and help heal by scar formation of the mucosal lining and regeneration of glandular mucosa and duodenal mucosal lining more rapidly. Inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis, are diseases which result in destruction of the mucosal surface of the small or large intestine, respectively. Thus, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to promote the resurfacing of the mucosal surface to aid more rapid healing and to prevent progression of inflammatory bowel disease. Treatment with polynucleotides or polypeptides, agonists or antagonists of the present invention, is expected to have a significant effect on the production of mucus throughout the gastrointestinal tract and could be used to protect the intestinal mucosa from injurious substances that are ingested or following surgery. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to treat diseases associate with the under expression.

Moreover, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to prevent and heal damage to the lungs due to various pathological states. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, which could stimulate proliferation and differentiation and promote the repair of alveoli and brochiolar epithelium to prevent or treat acute or chronic lung damage. For example, emphysema, which results in the progressive loss of aveoli, and inhalation injuries, i.e., resulting from smoke inhalation and burns, that cause necrosis of the bronchiolar epithelium and alveoli could be effectively treated using polynucleotides or polypeptides, agonists or antagonists of the present invention. Also, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to stimulate the proliferation of and differentiation of type II pneumocytes, which may help treat or prevent disease such as hyaline membrane diseases, such as infant respiratory distress syndrome and bronchopulmonary displasia, in premature infants.

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could stimulate the proliferation and differentiation of hepatocytes and, thus, could be used to alleviate or treat liver diseases and pathologies such as fulminant liver failure caused by cirrhosis, liver damage caused by viral hepatitis and toxic substances (i.e., acetaminophen, carbon tetraholoride and other hepatotoxins known in the art).

In addition, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used treat or prevent the onset of diabetes mellitus. In patients with newly diagnosed Types I and II diabetes, where some islet cell function remains, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used to maintain the islet function so as to alleviate, delay or prevent permanent manifestation of the disease. Also, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, could be used as an auxiliary in islet cell transplantation to improve or promote islet cell function.

Neural Activity and Neurological Diseases

The polynucleotides, polypeptides and agonists or antagonists of the invention may be used for the diagnosis and/or treatment of diseases, disorders, damage or injury of the brain and/or nervous system. Nervous system disorders that can be treated with the compositions of the invention (e.g., polypeptides, polynucleotides, and/or agonists or antagonists), include, but are not limited to, nervous system injuries, and diseases or disorders which result in either a disconnection of axons, a diminution or degeneration of neurons, or demyelination. Nervous system lesions which may be treated in a patient (including human and non-human mammalian patients) according to the methods of the invention, include but are not limited to, the following lesions of either the central (including spinal cord, brain) or peripheral nervous systems: (1) ischemic lesions, in which a lack of oxygen in a portion of the nervous system results in neuronal injury or death, including cerebral infarction or ischemia, or spinal cord infarction or ischemia; (2) traumatic lesions, including lesions caused by physical injury or associated with surgery, for example, lesions which sever a portion of the nervous system, or compression injuries; (3) malignant lesions, in which a portion of the nervous system is destroyed or injured by malignant tissue which is either a nervous system associated malignancy or a malignancy derived from non-nervous system tissue; (4) infectious lesions, in which a portion of the nervous system is destroyed or injured as a result of infection, for example, by an abscess or associated with infection by human immunodeficiency virus, herpes zoster, or herpes simplex virus or with Lyme disease, tuberculosis, or syphilis; (5) degenerative lesions, in which a portion of the nervous system is destroyed or injured as a result of a degenerative process including but not limited to, degeneration associated with Parkinson's disease, Alzheimer's disease, Huntington's chorea, or amyotrophic lateral sclerosis (ALS); (6) lesions associated with nutritional diseases or disorders, in which a portion of the nervous system is destroyed or injured by a nutritional disorder or disorder of metabolism including, but not limited to, vitamin B12 deficiency, folic acid deficiency, Wernicke disease, tobacco-alcohol amblyopia, Marchiafava-Bignami disease (primary degeneration of the corpus callosum), and alcoholic cerebellar degeneration; (7) neurological lesions associated with systemic diseases including, but not limited to, diabetes (diabetic neuropathy, Bell's palsy), systemic lupus erythematosus, carcinoma, or sarcoidosis; (8) lesions caused by toxic substances including alcohol, lead, or particular neurotoxins; and (9) demyelinated lesions in which a portion of the nervous system is destroyed or injured by a demyelinating disease including, but not limited to, multiple sclerosis, human immunodeficiency virus-associated myelopathy, transverse myelopathy or various etiologies, progressive multifocal leukoencephalopathy, and central pontine myelinolysis.

In one embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention are used to protect neural cells from the damaging effects of hypoxia. In a further preferred embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention are used to protect neural cells from the damaging effects of cerebral hypoxia. According to this embodiment, the compositions of the invention are used to treat or prevent neural cell injury associated with cerebral hypoxia. In one non-exclusive aspect of this embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention, are used to treat or prevent neural cell injury associated with cerebral ischemia. In another non-exclusive aspect of this embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention are used to treat or prevent neural cell injury associated with cerebral infarction.

In another preferred embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention are used to treat or prevent neural cell injury associated with a stroke. In a specific embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention are used to treat or prevent cerebral neural cell injury associated with a stroke.

In another preferred embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention are used to treat or prevent neural cell injury associated with a heart attack. In a specific embodiment, the polypeptides, polynucleotides, or agonists or antagonists of the invention are used to treat or prevent cerebral neural cell injury associated with a heart attack.

The compositions of the invention which are useful for treating or preventing a nervous system disorder may be selected by testing for biological activity in promoting the survival or differentiation of neurons. For example, and not by way of limitation, compositions of the invention which elicit any of the following effects may be useful according to the invention: (1) increased survival time of neurons in culture either in the presence or absence of hypoxia or hypoxic conditions; (2) increased sprouting of neurons in culture or in vivo; (3) increased production of a neuron-associated molecule in culture or in vivo, e.g., choline acetyltransferase or acetylcholinesterase with respect to motor neurons; or (4) decreased symptoms of neuron dysfunction in vivo. Such effects may be measured by any method known in the art. In preferred, non-limiting embodiments, increased survival of neurons may routinely be measured using a method set forth herein or otherwise known in the art, such as, for example, in Zhang et al., Proc Natl Acad Sci USA 97:3637-42 (2000) or in Arakawa et al., J. Neurosci., 10:3507-15 (1990); increased sprouting of neurons may be detected by methods known in the art, such as, for example, the methods set forth in Pestronk et al., Exp. Neurol., 70:65-82 (1980), or Brown et al., Ann. Rev. Neurosci., 4:17-42 (1981); increased production of neuron-associated molecules may be measured by bioassay, enzymatic assay, antibody binding, Northern blot assay, etc., using techniques known in the art and depending on the molecule to be measured; and motor neuron dysfunction may be measured by assessing the physical manifestation of motor neuron disorder, e.g., weakness, motor neuron conduction velocity, or functional disability.

In specific embodiments, motor neuron disorders that may be treated according to the invention include, but are not limited to, disorders such as infarction, infection, exposure to toxin, trauma, surgical damage, degenerative disease or malignancy that may affect motor neurons as well as other components of the nervous system, as well as disorders that selectively affect neurons such as amyotrophic lateral sclerosis, and including, but not limited to, progressive spinal muscular atrophy, progressive bulbar palsy, primary lateral sclerosis, infantile and juvenile muscular atrophy, progressive bulbar paralysis of childhood (Fazio-Londe syndrome), poliomyelitis and the post polio syndrome, and Hereditary Motorsensory Neuropathy (Charcot-Marie-Tooth Disease).

Further, polypeptides or polynucleotides of the invention may play a role in neuronal survival; synapse formation; conductance; neural differentiation, etc. Thus, compositions of the invention (including polynucleotides, polypeptides, and agonists or antagonists) may be used to diagnose and/or treat or prevent diseases or disorders associated with these roles, including, but not limited to, learning and/or cognition disorders. The compositions of the invention may also be useful in the treatment or prevention of neurodegenerative disease states and/or behavioural disorders. Such neurodegenerative disease states and/or behavioral disorders include, but are not limited to, Alzheimer's Disease, Parkinson's Disease, Huntington's Disease, Tourette Syndrome, schizophrenia, mania, dementia, paranoia, obsessive compulsive disorder, panic disorder, learning disabilities, ALS, psychoses, autism, and altered behaviors, including disorders in feeding, sleep patterns, balance, and perception. In addition, compositions of the invention may also play a role in the treatment, prevention and/or detection of developmental disorders associated with the developing embryo, or sexually-linked disorders.

Additionally, polypeptides, polynucleotides and/or agonists or antagonists of the invention, may be useful in protecting neural cells from diseases, damage, disorders, or injury, associated with cerebrovascular disorders including, but not limited to, carotid artery diseases (e.g., carotid artery thrombosis, carotid stenosis, or Moyamoya Disease), cerebral amyloid angiopathy, cerebral aneurysm, cerebral anoxia, cerebral arteriosclerosis, cerebral arteriovenous malformations, cerebral artery diseases, cerebral embolism and thrombosis (e.g., carotid artery thrombosis, sinus thrombosis, or Wallenberg's Syndrome), cerebral hemorrhage (e.g., epidural or subdural hematoma, or subarachnoid hemorrhage), cerebral infarction, cerebral ischemia (e.g., transient cerebral ischemia, Subclavian Steal Syndrome, or vertebrobasilar insufficiency), vascular dementia (e.g., multi-infarct), leukomalacia, periventricular, and vascular headache (e.g., cluster headache or migraines).

In accordance with yet a further aspect of the present invention, there is provided a process for utilizing polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, for therapeutic purposes, for example, to stimulate neurological cell proliferation and/or differentiation. Therefore, polynucleotides, polypeptides, agonists and/or antagonists of the invention may be used to treat and/or detect neurologic diseases. Moreover, polynucleotides or polypeptides, or agonists or antagonists of the invention, can be used as a marker or detector of a particular nervous system disease or disorder.

Examples of neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include brain diseases, such as metabolic brain diseases which includes phenylketonuria such as maternal phenylketonuria, pyruvate carboxylase deficiency, pyruvate dehydrogenase complex deficiency, Wernicke's Encephalopathy, brain edema, brain neoplasms such as cerebellar neoplasms which include infratentorial neoplasms, cerebral ventricle neoplasms such as choroid plexus neoplasms, hypothalamic neoplasms, supratentorial neoplasms, canavan disease, cerebellar diseases such as cerebellar ataxia which include spinocerebellar degeneration such as ataxia telangiectasia, cerebellar dyssynergia, Friederich's Ataxia, Machado-Joseph Disease, olivopontocerebellar atrophy, cerebellar neoplasms such as infratentorial neoplasms, diffuse cerebral sclerosis such as encephalitis periaxialis, globoid cell leukodystrophy, metachromatic leukodystrophy and subacute sclerosing panencephalitis.

Additional neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include cerebrovascular disorders (such as carotid artery diseases which include carotid artery thrombosis, carotid stenosis and Moyamoya Disease), cerebral amyloid angiopathy, cerebral aneurysm, cerebral anoxia, cerebral arteriosclerosis, cerebral arteriovenous malformations, cerebral artery diseases, cerebral embolism and thrombosis such as carotid artery thrombosis, sinus thrombosis and Wallenberg's Syndrome, cerebral hemorrhage such as epidural hematoma, subdural hematoma and subarachnoid hemorrhage, cerebral infarction, cerebral ischemia such as transient cerebral ischemia, Subclavian Steal Syndrome and vertebrobasilar insufficiency, vascular dementia such as multi-infarct dementia, periventricular leukomalacia, vascular headache such as cluster headache and migraine.

Additional neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include dementia such as AIDS Dementia Complex, presenile dementia such as Alzheimer's Disease and Creutzfeldt-Jakob Syndrome, senile dementia such as Alzheimer's Disease and progressive supranuclear palsy, vascular dementia such as multi-infarct dementia, encephalitis which include encephalitis periaxialis, viral encephalitis such as epidemic encephalitis, Japanese Encephalitis, St. Louis Encephalitis, tick-borne encephalitis and West Nile Fever, acute disseminated encephalomyelitis, meningoencephalitis such as uveomeningoencephalitic syndrome, Postencephalitic Parkinson Disease and subacute sclerosing panencephalitis, encephalomalacia such as periventricular leukomalacia, epilepsy such as generalized epilepsy which includes infantile spasms, absence epilepsy, myoclonic epilepsy which includes MERRF Syndrome, tonic-clonic epilepsy, partial epilepsy such as complex partial epilepsy, frontal lobe epilepsy and temporal lobe epilepsy, post-traumatic epilepsy, status epilepticus such as Epilepsia Partialis Continua, and Hallervorden-Spatz Syndrome.

Additional neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include hydrocephalus such as Dandy-Walker Syndrome and normal pressure hydrocephalus, hypothalamic diseases such as hypothalamic neoplasms, cerebral malaria, narcolepsy which includes cataplexy, bulbar poliomyelitis, cerebri pseudotumor, Rett Syndrome, Reye's Syndrome, thalamic diseases, cerebral toxoplasmosis, intracranial tuberculoma and Zellweger Syndrome, central nervous system infections such as AIDS Dementia Complex, Brain Abscess, subdural empyema, encephalomyelitis such as Equine Encephalomyelitis, Venezuelan Equine Encephalomyelitis, Necrotizing Hemorrhagic Encephalomyelitis, Visna, and cerebral malaria.

Additional neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include meningitis such as arachnoiditis, aseptic meningtitis such as viral meningtitis which includes lymphocytic choriomeningitis, Bacterial meningtitis which includes Haemophilus Meningtitis, Listeria Meningtitis, Meningococcal Meningtitis such as Waterhouse-Friderichsen Syndrome, Pneumococcal Meningtitis and meningeal tuberculosis, fungal meningitis such as Cryptococcal Meningtitis, subdural effusion, meningoencephalitis such as uvemeningoencephalitic syndrome, myelitis such as transverse myelitis, neurosyphilis such as tabes dorsalis, poliomyelitis which includes bulbar poliomyelitis and postpoliomyelitis syndrome, prion diseases (such as Creutzfeldt-Jakob Syndrome, Bovine Spongiform Encephalopathy, Gerstmann-Straussler Syndrome, Kuru, Scrapie), and cerebral toxoplasmosis.

Additional neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include central nervous system neoplasms such as brain neoplasms that include cerebellar neoplasms such as infratentorial neoplasms, cerebral ventricle neoplasms such as choroid plexus neoplasms, hypothalamic neoplasms and supratentorial neoplasms, meningeal neoplasms, spinal cord neoplasms which include epidural neoplasms, demyelinating diseases such as Canavan Diseases, diffuse cerebral sceloris which includes adrenoleukodystrophy, encephalitis periaxialis, globoid cell leukodystrophy, diffuse cerebral sclerosis such as metachromatic leukodystrophy, allergic encephalomyelitis, necrotizing hemorrhagic encephalomyelitis, progressive multifocal leukoencephalopathy, multiple sclerosis, central pontine myelinolysis, transverse myelitis, neuromyelitis optica, Scrapie, Swayback, Chronic Fatigue Syndrome, Visna, High Pressure Nervous Syndrome, Meningism, spinal cord diseases such as amyotonia congenita, amyotrophic lateral sclerosis, spinal muscular atrophy such as Werdnig-Hoffmann Disease, spinal cord compression, spinal cord neoplasms such as epidural neoplasms, syringomyelia, Tabes Dorsalis, Stiff-Man Syndrome, mental retardation such as Angelman Syndrome, Cri-du-Chat Syndrome, De Lange's Syndrome, Down Syndrome, Gangliosidoses such as gangliosidoses G(M1), Sandhoff Disease, Tay-Sachs Disease, Hartnup Disease, homocystinuria, Laurence-Moon-Biedl Syndrome, Lesch-Nyhan Syndrome, Maple Syrup Urine Disease, mucolipidosis such as fucosidosis, neuronal ceroid-lipofuscinosis, oculocerebrorenal syndrome, phenylketonuria such as maternal phenylketonuria, Prader-Willi Syndrome, Rett Syndrome, Rubinstein-Taybi Syndrome, Tuberous Sclerosis, WAGR Syndrome, nervous system abnormalities such as holoprosencephaly, neural tube defects such as anencephaly which includes hydrangencephaly, Arnold-Chairi Deformity, encephalocele, meningocele, meningomyelocele, spinal dysraphism such as spina bifida cystica and spina bifida occulta.

Additional neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include hereditary motor and sensory neuropathies which include Charcot-Marie Disease, Hereditary optic atrophy, Refsum's Disease, hereditary spastic paraplegia, Werdnig-Hoffmann Disease, Hereditary Sensory and Autonomic Neuropathies such as Congenital Analgesia and Familial Dysautonomia, Neurologic manifestations (such as agnosia that include Gerstmann's Syndrome, Amnesia such as retrograde amnesia, apraxia, neurogenic bladder, cataplexy, communicative disorders such as hearing disorders that includes deafness, partial hearing loss, loudness recruitment and tinnitus, language disorders such as aphasia which include agraphia, anomia, broca aphasia, and Wernicke Aphasia, Dyslexia such as Acquired Dyslexia, language development disorders, speech disorders such as aphasia which includes anomia, broca aphasia and Wernicke Aphasia, articulation disorders, communicative disorders such as speech disorders which include dysarthria, echolalia, mutism and stuttering, voice disorders such as aphonia and hoarseness, decerebrate state, delirium, fasciculation, hallucinations, meningism, movement disorders such as angelman syndrome, ataxia, athetosis, chorea, dystonia, hypokinesia, muscle hypotonia, myoclonus, tic, torticollis and tremor, muscle hypertonia such as muscle rigidity such as stiff-man syndrome, muscle spasticity, paralysis such as facial paralysis which includes Herpes Zoster Oticus, Gastroparesis, Hemiplegia, ophthalmoplegia such as diplopia, Duane's Syndrome, Horner's Syndrome, Chronic progressive external ophthalmoplegia such as Kearns Syndrome, Bulbar Paralysis, Tropical Spastic Paraparesis, Paraplegia such as Brown-Sequard Syndrome, quadriplegia, respiratory paralysis and vocal cord paralysis, paresis, phantom limb, taste disorders such as ageusia and dysgeusia, vision disorders such as amblyopia, blindness, color vision defects, diplopia, hemianopsia, scotoma and subnormal vision, sleep disorders such as hypersomnia which includes Kleine-Levin Syndrome, insomnia, and somnambulism, spasm such as trismus, unconsciousness such as coma, persistent vegetative state and syncope and vertigo, neuromuscular diseases such as amyotonia congenita, amyotrophic lateral sclerosis, Lambert-Eaton Myasthenic Syndrome, motor neuron disease, muscular atrophy such as spinal muscular atrophy, Charcot-Marie Disease and Werdnig-Hoffmann Disease, Postpoliomyelitis Syndrome, Muscular Dystrophy, Myasthenia Gravis, Myotonia Atrophica, Myotonia Confenita, Nemaline Myopathy, Familial Periodic Paralysis, Multiplex Paramyloclonus, Tropical Spastic Paraparesis and Stiff-Man Syndrome, peripheral nervous system diseases such as acrodynia, amyloid neuropathies, autonomic nervous system diseases such as Adie's Syndrome, Barre-Lieou Syndrome, Familial Dysautonomia, Horner's Syndrome, Reflex Sympathetic Dystrophy and Shy-Drager Syndrome, Cranial Nerve Diseases such as Acoustic Nerve Diseases such as Acoustic Neuroma which includes Neurofibromatosis 2, Facial Nerve Diseases such as Facial Neuralgia, Melkersson-Rosenthal Syndrome, ocular motility disorders which includes amblyopia, nystagmus, oculomotor nerve paralysis, ophthalmoplegia such as Duane's Syndrome, Horner's Syndrome, Chronic Progressive External Ophthalmoplegia which includes Kearns Syndrome, Strabismus such as Esotropia and Exotropia, Oculomotor Nerve Paralysis, Optic Nerve Diseases such as Optic Atrophy which includes Hereditary Optic Atrophy, Optic Disk Drusen, Optic Neuritis such as Neuromyelitis Optica, Papilledema, Trigeminal Neuralgia, Vocal Cord Paralysis, Demyelinating Diseases such as Neuromyelitis Optica and Swayback, and Diabetic neuropathies such as diabetic foot.

Additional neurologic diseases which can be treated or detected with polynucleotides, polypeptides, agonists, and/or antagonists of the present invention include nerve compression syndromes such as carpal tunnel syndrome, tarsal tunnel syndrome, thoracic outlet syndrome such as cervical rib syndrome, ulnar nerve compression syndrome, neuralgia such as causalgia, cervico-brachial neuralgia, facial neuralgia and trigeminal neuralgia, neuritis such as experimental allergic neuritis, optic neuritis, polyneuritis, polyradiculoneuritis and radiculities such as polyradiculitis, hereditary motor and sensory neuropathies such as Charcot-Marie Disease, Hereditary Optic Atrophy, Refsum's Disease, Hereditary Spastic Paraplegia and Werdnig-Hoffmann Disease, Hereditary Sensory and Autonomic Neuropathies which include Congenital Analgesia and Familial Dysautonomia, POEMS Syndrome, Sciatica, Gustatory Sweating and Tetany).

Endocrine Disorders

Polynucleotides or polypeptides, or agonists or antagonists of the present invention, may be used to treat, prevent, diagnose, and/or prognose disorders and/or diseases related to hormone imbalance, and/or disorders or diseases of the endocrine system.

Hormones secreted by the glands of the endocrine system control physical growth, sexual function, metabolism, and other functions. Disorders may be classified in two ways: disturbances in the production of hormones, and the inability of tissues to respond to hormones. The etiology of these hormone imbalance or endocrine system diseases, disorders or conditions may be genetic, somatic, such as cancer and some autoimmune diseases, acquired (e.g., by chemotherapy, injury or toxins), or infectious. Moreover, polynucleotides, polypeptides, antibodies, and/or agonists or antagonists of the present invention can be used as a marker or detector of a particular disease or disorder related to the endocrine system and/or hormone imbalance.

Endocrine system and/or hormone imbalance and/or diseases encompass disorders of uterine motility including, but not limited to: complications with pregnancy and labor (e.g., pre-term labor, post-term pregnancy, spontaneous abortion, and slow or stopped labor); and disorders and/or diseases of the menstrual cycle (e.g., dysmenorrhea and endometriosis).

Endocrine system and/or hormone imbalance disorders and/or diseases include disorders and/or diseases of the pancreas, such as, for example, diabetes mellitus, diabetes insipidus, congenital pancreatic agenesis, pheochromocytoma—islet cell tumor syndrome; disorders and/or diseases of the adrenal glands such as, for example, Addison's Disease, corticosteroid deficiency, virilizing disease, hirsutism, Cushing's Syndrome, hyperaldosteronism, pheochromocytoma; disorders and/or diseases of the pituitary gland, such as, for example, hyperpituitarism, hypopituitarism, pituitary dwarfism, pituitary adenoma, panhypopituitarism, acromegaly, gigantism; disorders and/or diseases of the thyroid, including but not limited to, hyperthyroidism, hypothyroidism, Plummer's disease, Graves' disease (toxic diffuse goiter), toxic nodular goiter, thyroiditis (Hashimoto's thyroiditis, subacute granulomatous thyroiditis, and silent lymphocytic thyroiditis), Pendred's syndrome, myxedema, cretinism, thyrotoxicosis, thyroid hormone coupling defect, thymic aplasia, Hurthle cell tumours of the thyroid, thyroid cancer, thyroid carcinoma, Medullary thyroid carcinoma; disorders and/or diseases of the parathyroid, such as, for example, hyperparathyroidism, hypoparathyroidism; disorders and/or diseases of the hypothalamus.

In addition, endocrine system and/or hormone imbalance disorders and/or diseases may also include disorders and/or diseases of the testes or ovaries, including cancer. Other disorders and/or diseases of the testes or ovaries further include, for example, ovarian cancer, polycystic ovary syndrome, Klinefelter's syndrome, vanishing testes syndrome (bilateral anorchia), congenital absence of Leydig's cells, cryptorchidism, Noonan's syndrome, myotonic dystrophy, capillary haemangioma of the testis (benign), neoplasias of the testis and neo-testis.

Moreover, endocrine system and/or hormone imbalance disorders and/or diseases may also include disorders and/or diseases such as, for example, polyglandular deficiency syndromes, pheochromocytoma, neuroblastoma, multiple Endocrine neoplasia, and disorders and/or cancers of endocrine tissues.

In another embodiment, a polypeptide of the invention, or polynucleotides, antibodies, agonists, or antagonists corresponding to that polypeptide, may be used to diagnose, prognose, prevent, and/or treat endocrine diseases and/or disorders associated with the tissue(s) in which the polypeptide of the invention is expressed, including one, two, three, four, five, or more tissues disclosed in Table 1B.2, column 5 (Tissue Distribution Library Code).

Reproductive System Disorders

The polynucleotides or polypeptides, or agonists or antagonists of the invention may be used for the diagnosis, treatment, or prevention of diseases and/or disorders of the reproductive system. Reproductive system disorders that can be treated by the compositions of the invention, include, but are not limited to, reproductive system injuries, infections, neoplastic disorders, congenital defects, and diseases or disorders which result in infertility, complications with pregnancy, labor, or parturition, and postpartum difficulties.

Reproductive system disorders and/or diseases include diseases and/or disorders of the testes, including testicular atrophy, testicular feminization, cryptorchism (unilateral and bilateral), anorchia, ectopic testis, epididymitis and orchitis (typically resulting from infections such as, for example, gonorrhea, mumps, tuberculosis, and syphilis), testicular torsion, vasitis nodosa, germ cell tumors (e.g., seminomas, embryonal cell carcinomas, teratocarcinomas, choriocarcinomas, yolk sac tumors, and teratomas), stromal tumors (e.g., Leydig cell tumors), hydrocele, hematocele, varicocele, spermatocele, inguinal hernia, and disorders of sperm production (e.g., immotile cilia syndrome, aspermia, asthenozoospermia, azoospermia, oligospermia, and teratozoospermia).

Reproductive system disorders also include disorders of the prostate gland, such as acute non-bacterial prostatitis, chronic non-bacterial prostatitis, acute bacterial prostatitis, chronic bacterial prostatitis, prostatodystonia, prostatosis, granulomatous prostatitis, malacoplakia, benign prostatic hypertrophy or hyperplasia, and prostate neoplastic disorders, including adenocarcinomas, transitional cell carcinomas, ductal carcinomas, and squamous cell carcinomas.

Additionally, the compositions of the invention may be useful in the diagnosis, treatment, and/or prevention of disorders or diseases of the penis and urethra, including inflammatory disorders, such as balanoposthitis, balanitis xerotica obliterans, phimosis, paraphimosis, syphilis, herpes simplex virus, gonorrhea, non-gonococcal urethritis, chlamydia, mycoplasma, trichomonas, HIV, AIDS, Reiter's syndrome, condyloma acuminatum, condyloma latum, and pearly penile papules; urethral abnormalities, such as hypospadias, epispadias, and phimosis; premalignant lesions, including Erythroplasia of Queyrat, Bowen's disease, Bowenoid paplosis, giant condyloma of Buscke-Lowenstein, and varrucous carcinoma; penile cancers, including squamous cell carcinomas, carcinoma in situ, verrucous carcinoma, and disseminated penile carcinoma; urethral neoplastic disorders, including penile urethral carcinoma, bulbomembranous urethral carcinoma, and prostatic urethral carcinoma; and erectile disorders, such as priapism, Peyronie's disease, erectile dysfunction, and impotence.

Moreover, diseases and/or disorders of the vas deferens include vasculititis and CBAVD (congenital bilateral absence of the vas deferens); additionally, the polynucleotides, polypeptides, and agonists or antagonists of the present invention may be used in the diagnosis, treatment, and/or prevention of diseases and/or disorders of the seminal vesicles, including hydatid disease, congenital chloride diarrhea, and polycystic kidney disease.

Other disorders and/or diseases of the male reproductive system include, for example, Klinefelter's syndrome, Young's syndrome, premature ejaculation, diabetes mellitus, cystic fibrosis, Kartagener's syndrome, high fever, multiple sclerosis, and gynecomastia.

Further, the polynucleotides, polypeptides, and agonists or antagonists of the present invention may be used in the diagnosis, treatment, and/or prevention of diseases and/or disorders of the vagina and vulva, including bacterial vaginosis, candida vaginitis, herpes simplex virus, chancroid, granuloma inguinale, lymphogranuloma venereum, scabies, human papillomavirus, vaginal trauma, vulvar trauma, adenosis, chiamydia vaginitis, gonorrhea, trichomonas vaginitis, condyloma acuminatum, syphilis, molluscum contagiosum, atrophic vaginitis, Paget's disease, lichen sclerosus, lichen planus, vulvodynia, toxic shock syndrome, vaginismus, vulvovaginitis, vulvar vestibulitis, and neoplastic disorders, such as squamous cell hyperplasia, clear cell carcinoma, basal cell carcinoma, melanomas, cancer of Bartholin's gland, and vulvar intraepithelial neoplasia.

Disorders and/or diseases of the uterus include dysmenorrhea, retroverted uterus, endometriosis, fibroids, adenomyosis, anovulatory bleeding, amenorrhea, Cushing's syndrome, hydatidiform moles, Asherman's syndrome, premature menopause, precocious puberty, uterine polyps, dysfunctional uterine bleeding (e.g., due to aberrant hormonal signals), and neoplastic disorders, such as adenocarcinomas, keiomyosarcomas, and sarcomas. Additionally, the polypeptides, polynucleotides, or agonists or antagonists of the invention may be useful as a marker or detector of, as well as in the diagnosis, treatment, and/or prevention of congenital uterine abnormalities, such as bicornuate uterus, septate uterus, simple unicornuate uterus, unicornuate uterus with a noncavitary rudimentary horn, unicornuate uterus with a non-communicating cavitary rudimentary horn, unicornuate uterus with a communicating cavitary horn, arcuate uterus, uterine didelfus, and T-shaped uterus.

Ovarian diseases and/or disorders include anovulation, polycystic ovary syndrome (Stein-Leventhal syndrome), ovarian cysts, ovarian hypofunction, ovarian insensitivity to gonadotropins, ovarian overproduction of androgens, right ovarian vein syndrome, amenorrhea, hirutism, and ovarian cancer (including, but not limited to, primary and secondary cancerous growth, Sertoli-Leydig tumors, endometriod carcinoma of the ovary, ovarian papillary serous adenocarcinoma, ovarian mucinous adenocarcinoma, and Ovarian Krukenberg tumors).

Cervical diseases and/or disorders include cervicitis, chronic cervicitis, mucopurulent cervicitis, cervical dysplasia, cervical polyps, Nabothian cysts, cervical erosion, cervical incompetence, and cervical neoplasms (including, for example, cervical carcinoma, squamous metaplasia, squamous cell carcinoma, adenosquamous cell neoplasia, and columnar cell neoplasia).

Additionally, diseases and/or disorders of the reproductive system include disorders and/or diseases of pregnancy, including miscarriage and stillbirth, such as early abortion, late abortion, spontaneous abortion, induced abortion, therapeutic abortion, threatened abortion, missed abortion, incomplete abortion, complete abortion, habitual abortion, missed abortion, and septic abortion; ectopic pregnancy, anemia, Rh incompatibility, vaginal bleeding during pregnancy, gestational diabetes, intrauterine growth retardation, polyhydramnios, HELLP syndrome, abruptio placentae, placenta previa, hyperemesis, preeclampsia, eclampsia, herpes gestationis, and urticaria of pregnancy. Additionally, the polynucleotides, polypeptides, and agonists or antagonists of the present invention may be used in the diagnosis, treatment, and/or prevention of diseases that can complicate pregnancy, including heart disease, heart failure, rheumatic heart disease, congenital heart disease, mitral valve prolapse, high blood pressure, anemia, kidney disease, infectious disease (e.g., rubella, cytomegalovirus, toxoplasmosis, infectious hepatitis, chlamydia, HIV, AIDS, and genital herpes), diabetes mellitus, Graves' disease, thyroiditis, hypothyroidism, Hashimoto's thyroiditis, chronic active hepatitis, cirrhosis of the liver, primary biliary cirrhosis, asthma, systemic lupus eryematosis, rheumatoid arthritis, myasthenia gravis, idiopathic thrombocytopenic purpura, appendicitis, ovarian cysts, gallbladder disorders, and obstruction of the intestine.

Complications associated with labor and parturition include premature rupture of the membranes, pre-term labor, post-term pregnancy, postmaturity, labor that progresses too slowly, fetal distress (e.g., abnormal heart rate (fetal or maternal), breathing problems, and abnormal fetal position), shoulder dystocia, prolapsed umbilical cord, amniotic fluid embolism, and aberrant uterine bleeding.

Further, diseases and/or disorders of the postdelivery period, including endometritis, myometritis, parametritis, peritonitis, pelvic thrombophlebitis, pulmonary embolism, endotoxemia, pyelonephritis, saphenous thrombophlebitis, mastitis, cystitis, postpartum hemorrhage, and inverted uterus.

Other disorders and/or diseases of the female reproductive system that may be diagnosed, treated, and/or prevented by the polynucleotides, polypeptides, and agonists or antagonists of the present invention include, for example, Turner's syndrome, pseudohermaphroditism, premenstrual syndrome, pelvic inflammatory disease, pelvic congestion (vascular engorgement), frigidity, anorgasmia, dyspareunia, ruptured fallopian tube, and Mittelschmerz.

Infectious Disease

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention can be used to treat or detect infectious agents. For example, by increasing the immune response, particularly increasing the proliferation and differentiation of B and/or T cells, infectious diseases may be treated. The immune response may be increased by either enhancing an existing immune response, or by initiating a new immune response. Alternatively, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention may also directly inhibit the infectious agent, without necessarily eliciting an immune response.

Viruses are one example of an infectious agent that can cause disease or symptoms that can be treated or detected by a polynucleotide or polypeptide and/or agonist or antagonist of the present invention. Examples of viruses, include, but are not limited to Examples of viruses, include, but are not limited to the following DNA and RNA viruses and viral families: Arbovirus, Adenoviridae, Arenaviridae, Arterivirus, Birnaviridae, Bunyaviridae, Caliciviridae, Circoviridae, Coronaviridae, Dengue, EBV, HIV, Flaviviridae, Hepadnaviridae (Hepatitis), Herpesviridae (such as, Cytomegalovirus, Herpes Simplex, Herpes Zoster), Mononegavirus (e.g., Paramyxoviridae, Morbillivirus, Rhabdoviridae), Orthomyxoviridae (e.g., Influenza A, Influenza B, and parainfluenza), Papiloma virus, Papovaviridae, Parvoviridae, Picornaviridae, Poxyiridae (such as Smallpox or Vaccinia), Reoviridae (e.g., Rotavirus), Retroviridae (HTLV-I, HTLV-II, Lentivirus), and Togaviridae (e.g., Rubivirus). Viruses falling within these families can cause a variety of diseases or symptoms, including, but not limited to: arthritis, bronchiollitis, respiratory syncytial virus, encephalitis, eye infections (e.g., conjunctivitis, keratitis), chronic fatigue syndrome, hepatitis (A, B, C, E, Chronic Active, Delta), Japanese B encephalitis, Junin, Chikungunya, Rift Valley fever, yellow fever, meningitis, opportunistic infections (e.g., AIDS), pneumonia, Burkitt's Lymphoma, chickenpox, hemorrhagic fever, Measles, Mumps, Parainfluenza, Rabies, the common cold, Polio, leukemia, Rubella, sexually transmitted diseases, skin diseases (e.g., Kaposi's, warts), and viremia. polynucleotides or polypeptides, or agonists or antagonists of the invention, can be used to treat or detect any of these symptoms or diseases. In specific embodiments, polynucleotides, polypeptides, or agonists or antagonists of the invention are used to treat: meningitis, Dengue, EBV, and/or hepatitis (e.g., hepatitis B). In an additional specific embodiment polynucleotides, polypeptides, or agonists or antagonists of the invention are used to treat patients nonresponsive to one or more other commercially available hepatitis vaccines. In a further specific embodiment polynucleotides, polypeptides, or agonists or antagonists of the invention are used to treat AIDS.

Similarly, bacterial and fungal agents that can cause disease or symptoms and that can be treated or detected by a polynucleotide or polypeptide and/or agonist or antagonist of the present invention include, but not limited to, the following Gram-Negative and Gram-positive bacteria, bacterial families, and fungi: Actinomyces (e.g., Norcardia), Acinetobacter, Cryptococcus neoformans, Aspergillus, Bacillaceae (e.g., Bacillus anthrasis), Bacteroides (e.g., Bacteroides fragilis), Blastomycosis, Bordetella, Borrelia (e.g., Borrelia burgdorferi), Brucella, Candidia, Campylobacter, Chlamydia, Clostridium (e.g., Clostridium botulinum, Clostridium dificile, Clostridium perfringens, Clostridium tetani), Coccidioides, Corynebacterium (e.g., Corynebacterium diptheriae), Cryptococcus, Dermatocycoses, E. coli (e.g., Enterotoxigenic E. coli and Enterohemorrhagic E. coli), Enterobacter (e.g. Enterobacter aerogenes), Enterobacteriaceae (Klebsiella, Salmonella (e.g., Salmonella typhi, Salmonella enteritidis, Salmonella typhi), Serratia, Yersinia, Shigella), Erysipelothrix, Haemophilus (e.g., Haemophilus influenza type B), Helicobacter, Legionella (e.g., Legionella pneumophila), Leptospira, Listeria (e.g., Listeria monocytogenes), Mycoplasma, Mycobacterium (e.g., Mycobacterium leprae and Mycobacterium tuberculosis), Vibrio (e.g., Vibrio cholerae), Neisseriaceae (e.g., Neisseria gonorrhea, Neisseria meningitidis), Pasteurellacea, Proteus, Pseudomonas (e.g., Pseudomonas aeruginosa), Rickettsiaceae, Spirochetes (e.g., Treponema spp., Leptospira spp., Borrelia spp.), Shigella spp., Staphylococcus (e.g., Staphylococcus aureus), Meningiococcus, Pneumococcus and Streptococcus (e.g., Streptococcus pneumoniae and Groups A, B, and C Streptococci), and Ureaplasmas. These bacterial, parasitic, and fungal families can cause diseases or symptoms, including, but not limited to: antibiotic-resistant infections, bacteremia, endocarditis, septicemia, eye infections (e.g., conjunctivitis), uveitis, tuberculosis, gingivitis, bacterial diarrhea, opportunistic infections (e.g., AIDS related infections), paronychia, prosthesis-related infections, dental caries, Reiter's Disease, respiratory tract infections, such as Whooping Cough or Empyema, sepsis, Lyme Disease, Cat-Scratch Disease, dysentery, paratyphoid fever, food poisoning, Legionella disease, chronic and acute inflammation, erythema, yeast infections, typhoid, pneumonia, gonorrhea, meningitis (e.g., mengitis types A and B), chiamydia, syphillis, diphtheria, leprosy, brucellosis, peptic ulcers, anthrax, spontaneous abortions, birth defects, pneumonia, lung infections, ear infections, deafness, blindness, lethargy, malaise, vomiting, chronic diarrhea, Crohn's disease, colitis, vaginosis, sterility, pelvic inflammatory diseases, candidiasis, paratuberculosis, tuberculosis, lupus, botulism, gangrene, tetanus, impetigo, Rheumatic Fever, Scarlet Fever, sexually transmitted diseases, skin diseases (e.g., cellulitis, dermatocycoses), toxemia, urinary tract infections, wound infections, noscomial infections. Polynucleotides or polypeptides, agonists or antagonists of the invention, can be used to treat or detect any of these symptoms or diseases. In specific embodiments, polynucleotides, polypeptides, agonists or antagonists of the invention are used to treat: tetanus, diptheria, botulism, and/or meningitis type B.

Moreover, parasitic agents causing disease or symptoms that can be treated, prevented, and/or diagnosed by a polynucleotide or polypeptide and/or agonist or antagonist of the present invention include, but not limited to, the following families or class: Amebiasis, Babesiosis, Coccidiosis, Cryptosporidiosis, Dientamoebiasis, Dourine, Ectoparasitic, Giardias, Helminthiasis, Leishmaniasis, Schistisoma, Theileriasis, Toxoplasmosis, Trypanosomiasis, and Trichomonas and Sporozoans (e.g., Plasmodium virax, Plasmodium falciparium, Plasmodium malariae and Plasmodium ovale). These parasites can cause a variety of diseases or symptoms, including, but not limited to: Scabies, Trombiculiasis, eye infections, intestinal disease (e.g., dysentery, giardiasis), liver disease, lung disease, opportunistic infections (e.g., AIDS related), malaria, pregnancy complications, and toxoplasmosis. polynucleotides or polypeptides, or agonists or antagonists of the invention, can be used to treat, prevent, and/or diagnose any of these symptoms or diseases. In specific embodiments, polynucleotides, polypeptides, or agonists or antagonists of the invention are used to treat, prevent, and/or diagnose malaria.

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention of the present invention could either be by administering an effective amount of a polypeptide to the patient, or by removing cells from the patient, supplying the cells with a polynucleotide of the present invention, and returning the engineered cells to the patient (ex vivo therapy). Moreover, the polypeptide or polynucleotide of the present invention can be used as an antigen in a vaccine to raise an immune response against infectious disease.

Regeneration

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention can be used to differentiate, proliferate, and attract cells, leading to the regeneration of tissues. (See, Science 276:59-87 (1997)). The regeneration of tissues could be used to repair, replace, or protect tissue damaged by congenital defects, trauma (wounds, burns, incisions, or ulcers), age, disease (e.g. osteoporosis, osteocarthritis, periodontal disease, liver failure), surgery, including cosmetic plastic surgery, fibrosis, reperfusion injury, or systemic cytokine damage.

Tissues that could be regenerated using the present invention include organs (e.g., pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth, skeletal or cardiac), vasculature (including vascular and lymphatics), nervous, hematopoietic, and skeletal (bone, cartilage, tendon, and ligament) tissue. Preferably, regeneration occurs without or decreased scarring. Regeneration also may include angiogenesis.

Moreover, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, may increase regeneration of tissues difficult to heal. For example, increased tendon/ligament regeneration would quicken recovery time after damage. Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention could also be used prophylactically in an effort to avoid damage. Specific diseases that could be treated include of tendinitis, carpal tunnel syndrome, and other tendon or ligament defects. A further example of tissue regeneration of non-healing wounds includes pressure ulcers, ulcers associated with vascular insufficiency, surgical, and traumatic wounds.

Similarly, nerve and brain tissue could also be regenerated by using polynucleotides or polypeptides, as well as agonists or antagonists of the present invention, to proliferate and differentiate nerve cells. Diseases that could be treated using this method include central and peripheral nervous system diseases, neuropathies, or mechanical and traumatic disorders (e.g., spinal cord disorders, head trauma, cerebrovascular disease, and stoke). Specifically, diseases associated with peripheral nerve injuries, peripheral neuropathy (e.g., resulting from chemotherapy or other medical therapies), localized neuropathies, and central nervous system diseases (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Shy-Drager syndrome), could all be treated using the polynucleotides or polypeptides, as well as agonists or antagonists of the present invention.

Gastrointestinal Disorders

Polynucleotides or polypeptides, or agonists or antagonists of the present invention, may be used to treat, prevent, diagnose, and/or prognose gastrointestinal disorders, including inflammatory diseases and/or conditions, infections, cancers (e.g., intestinal neoplasms (carcinoid tumor of the small intestine, non-Hodgkin's lymphoma of the small intestine, small bowl lymphoma)), and ulcers, such as peptic ulcers.

Gastrointestinal disorders include dysphagia, odynophagia, inflammation of the esophagus, peptic esophagitis, gastric reflux, submucosal fibrosis and stricturing, Mallory-Weiss lesions, leiomyomas, lipomas, epidermal cancers, adeoncarcinomas, gastric retention disorders, gastroenteritis, gastric atrophy, gastric/stomach cancers, polyps of the stomach, autoimmune disorders such as pernicious anemia, pyloric stenosis, gastritis (bacterial, viral, eosinophilic, stress-induced, chronic erosive, atrophic, plasma cell, and Ménétrier's), and peritoneal diseases (e.g., chyloperioneum, hemoperitoneum, mesenteric cyst, mesenteric lymphadenitis, mesenteric vascular occlusion, panniculitis, neoplasms, peritonitis, pneumoperitoneum, bubphrenic abscess,).

Gastrointestinal disorders also include disorders associated with the small intestine, such as malabsorption syndromes, distension, irritable bowel syndrome, sugar intolerance, celiac disease, duodenal ulcers, duodenitis, tropical sprue, Whipple's disease, intestinal lymphangiectasia, Crohn's disease, appendicitis, obstructions of the ileum, Meckel's diverticulum, multiple diverticula, failure of complete rotation of the small and large intestine, lymphoma, and bacterial and parasitic diseases (such as Traveler's diarrhea, typhoid and paratyphoid, cholera, infection by Roundworms (Ascariasis lumbricoides), Hookworms (Ancylostoma duodenale), Threadworms (Enterobius vermicularis), Tapeworms (Taenia saginata, Echinococcus granulosus, Diphyllobothrium spp., and T. solium).

Liver diseases and/or disorders include intrahepatic cholestasis (alagille syndrome, biliary liver cirrhosis), fatty liver (alcoholic fatty liver, reye syndrome), hepatic vein thrombosis, hepatolentricular degeneration, hepatomegaly, hepatopulmonary syndrome, hepatorenal syndrome, portal hypertension (esophageal and gastric varices), liver abscess (amebic liver abscess), liver cirrhosis (alcoholic, biliary and experimental), alcoholic liver diseases (fatty liver, hepatitis, cirrhosis), parasitic (hepatic echinococcosis, fascioliasis, amebic liver abscess), jaundice (hemolytic, hepatocellular, and cholestatic), cholestasis, portal hypertension, liver enlargement, ascites, hepatitis (alcoholic hepatitis, animal hepatitis, chronic hepatitis (autoimmune, hepatitis B, hepatitis C, hepatitis D, drug induced), toxic hepatitis, viral human hepatitis (hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E), Wilson's disease, granulomatous hepatitis, secondary biliary cirrhosis, hepatic encephalopathy, portal hypertension, varices, hepatic encephalopathy, primary biliary cirrhosis, primary sclerosing cholangitis, hepatocellular adenoma, hemangiomas, bile stones, liver failure (hepatic encephalopathy, acute liver failure), and liver neoplasms (angiomyolipoma, calcified liver metastases, cystic liver metastases, epithelial tumors, fibrolamellar hepatocarcinoma, focal nodular hyperplasia, hepatic adenoma, hepatobiliary cystadenoma, hepatoblastoma, hepatocellular carcinoma, hepatoma, liver cancer, liver hemangioendothelioma, mesenchymal hamartoma, mesenchymal tumors of liver, nodular regenerative hyperplasia, benign liver tumors (Hepatic cysts [Simple cysts, Polycystic liver disease, Hepatobiliary cystadenoma, Choledochal cyst], Mesenchymal tumors [Mesenchymal hamartoma, Infantile hemangioendothelioma, Hemangioma, Peliosis hepatis, Lipomas, Inflammatory pseudotumor, Miscellaneous], Epithelial tumors [Bile duct epithelium (Bile duct hamartoma, Bile duct adenoma), Hepatocyte (Adenoma, Focal nodular hyperplasia, Nodular regenerative hyperplasia)], malignant liver tumors [hepatocellular, hepatoblastoma, hepatocellular carcinoma, cholangiocellular, cholangiocarcinoma, cystadenocarcinoma, tumors of blood vessels, angiosarcoma, Karposi's sarcoma, hemangioendothelioma, other tumors, embryonal sarcoma, fibrosarcoma, leiomyosarcoma, rhabdomyosarcoma, carcinosarcoma, teratoma, carcinoid, squamous carcinoma, primary lymphomal), peliosis hepatis, erythrohepatic porphyria, hepatic porphyria (acute intermittent porphyria, porphyria cutanea tarda), Zellweger syndrome).

Pancreatic diseases and/or disorders include acute pancreatitis, chronic pancreatitis (acute necrotizing pancreatitis, alcoholic pancreatitis), neoplasms (adenocarcinoma of the pancreas, cystadenocarcinoma, insulinoma, gastrinoma, and glucagonoma, cystic neoplasms, islet-cell tumors, pancreoblastoma), and other pancreatic diseases (e.g., cystic, fibrosis, cyst (pancreatic pseudocyst, pancreatic fistula, insufficiency)).

Gallbladder diseases include gallstones (cholelithiasis and choledocholithiasis), postcholecystectomy syndrome, diverticulosis of the gallbladder, acute cholecystitis, chronic cholecystitis, bile duct tumors, and mucocele.

Diseases and/or disorders of the large intestine include antibiotic-associated colitis, diverticulitis, ulcerative colitis, acquired megacolon, abscesses, fungal and bacterial infections, anorectal disorders (e.g., fissures, hemorrhoids), colonic diseases (colitis, colonic neoplasms [colon cancer, adenomatous colon polyps (e.g., villous adenoma), colon carcinoma, colorectal cancer], colonic diverticulitis, colonic diverticulosis, megacolon [Hirschsprung disease, toxic megacolon]; sigmoid diseases [proctocolitis, sigmoin neoplasms]), constipation, Crohn's disease, diarrhea (infantile diarrhea, dysentery), duodenal diseases (duodenal neoplasms, duodenal obstruction, duodenal ulcer, duodenitis), enteritis (enterocolitis), HIV enteropathy, ileal diseases (ileal neoplasms, ileitis), immunoproliferative small intestinal disease, inflammatory bowel disease (ulcerative colitis, Crohn's disease), intestinal atresia, parasitic diseases (anisakiasis, balantidiasis, blastocystis infections, cryptosporidiosis, dientamoebiasis, amebic dysentery, giardiasis), intestinal fistula (rectal fistula), intestinal neoplasms (cecal neoplasms, colonic neoplasms, duodenal neoplasms, ileal neoplasms, intestinal polyps, jejunal neoplasms, rectal neoplasms), intestinal obstruction (afferent loop syndrome, duodenal obstruction, impacted feces, intestinal pseudo-obstruction [cecal volvulus], intussusception), intestinal perforation, intestinal polyps (colonic polyps, gardner syndrome, peutz-jeghers syndrome), jejunal diseases (jejunal neoplasms), malabsorption syndromes (blind loop syndrome, celiac disease, lactose intolerance, short bowl syndrome, tropical sprue, whipple's disease), mesenteric vascular occlusion, pneumatosis cystoides intestinalis, protein-losing enteropathies (intestinal lymphagiectasis), rectal diseases (anus diseases, fecal incontinence, hemorrhoids, proctitis, rectal fistula, rectal prolapse, rectocele), peptic ulcer (duodenal ulcer, peptic esophagitis, hemorrhage, perforation, stomach ulcer, Zollinger-Ellison syndrome), postgastrectomy syndromes (dumping syndrome), stomach diseases (e.g., achlorhydria, duodenogastric reflux (bile reflux), gastric antral vascular ectasia, gastric fistula, gastric outlet obstruction, gastritis (atrophic or hypertrophic), gastroparesis, stomach dilatation, stomach diverticulum, stomach neoplasms (gastric cancer, gastric polyps, gastric adenocarcinoma, hyperplastic gastric polyp), stomach rupture, stomach ulcer, stomach volvulus), tuberculosis, visceroptosis, vomiting (e.g., hematemesis, hyperemesis gravidarum, postoperative nausea and vomiting) and hemorrhagic colitis.

Further diseases and/or disorders of the gastrointestinal system include biliary tract diseases, such as, gastroschisis, fistula (e.g., biliary fistula, esophageal fistula, gastric fistula, intestinal fistula, pancreatic fistula), neoplasms (e.g., biliary tract neoplasms, esophageal neoplasms, such as adenocarcinoma of the esophagus, esophageal squamous cell carcinoma, gastrointestinal neoplasms, pancreatic neoplasms, such as adenocarcinoma of the pancreas, mucinous cystic neoplasm of the pancreas, pancreatic cystic neoplasms, pancreatoblastoma, and peritoneal neoplasms), esophageal disease (e.g., bullous diseases, candidiasis, glycogenic acanthosis, ulceration, barrett esophagus varices, atresia, cyst, diverticulum (e.g., Zenker's diverticulum), fistula (e.g., tracheoesophageal fistula), motility disorders (e.g., CREST syndrome, deglutition disorders, achalasia, spasm, gastroesophageal reflux), neoplasms, perforation (e.g., Boerhaave syndrome, Mallory-Weiss syndrome), stenosis, esophagitis, diaphragmatic hernia (e.g., hiatal hernia); gastrointestinal diseases, such as, gastroenteritis (e.g., cholera morbus, norwalk virus infection), hemorrhage (e.g., hematemesis, melena, peptic ulcer hemorrhage), stomach neoplasms (gastric cancer, gastric polyps, gastric adenocarcinoma, stomach cancer)), hernia (e.g., congenital diaphragmatic hernia, femoral hernia, inguinal hernia, obturator hernia, umbilical hernia, ventral hernia), and intestinal diseases (e.g., cecal diseases (appendicitis, cecal neoplasms)).

Chemotaxis

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention may have chemotaxis activity. A chemotaxic molecule attracts or mobilizes cells (e.g., monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells) to a particular site in the body, such as inflammation, infection, or site of hyperproliferation. The mobilized cells can then fight off and/or heal the particular trauma or abnormality.

Polynucleotides or polypeptides, as well as agonists or antagonists of the present invention may increase chemotaxic activity of particular cells. These chemotactic molecules can then be used to treat inflammation, infection, hyperproliferative disorders, or any immune system disorder by increasing the number of cells targeted to a particular location in the body. For example, chemotaxic molecules can be used to treat wounds and other trauma to tissues by attracting immune cells to the injured location. Chemotactic molecules of the present invention can also attract fibroblasts, which can be used to treat wounds.

It is also contemplated that polynucleotides or polypeptides, as well as agonists or antagonists of the present invention may inhibit chemotactic activity. These molecules could also be used to treat disorders. Thus, polynucleotides or polypeptides, as well as agonists or antagonists of the present invention could be used as an inhibitor of chemotaxis.

Binding Activity

A polypeptide of the present invention may be used to screen for molecules that bind to the polypeptide or for molecules to which the polypeptide binds. The binding of the polypeptide and the molecule may activate (agonist), increase, inhibit (antagonist), or decrease activity of the polypeptide or the molecule bound. Examples of such molecules include antibodies, oligonucleotides, proteins (e.g., receptors), or small molecules.

Preferably, the molecule is closely related to the natural ligand of the polypeptide, e.g., a fragment of the ligand, or a natural substrate, a ligand, a structural or functional mimetic. (See, Coligan et al., Current Protocols in Immunology 1(2):Chapter 5 (1991)). Similarly, the molecule can be closely related to the natural receptor to which the polypeptide binds, or at least, a fragment of the receptor capable of being bound by the polypeptide (e.g., active site). In either case, the molecule can be rationally designed using known techniques.

Preferably, the screening for these molecules involves producing appropriate cells which express the polypeptide. Preferred cells include cells from mammals, yeast, Drosophila, or E. coli. Cells expressing the polypeptide (or cell membrane containing the expressed polypeptide) are then preferably contacted with a test compound potentially containing the molecule to observe binding, stimulation, or inhibition of activity of either the polypeptide or the molecule.

The assay may simply test binding of a candidate compound to the polypeptide, wherein binding is detected by a label, or in an assay involving competition with a labeled competitor. Further, the assay may test whether the candidate compound results in a signal generated by binding to the polypeptide.

Alternatively, the assay can be carried out using cell-free preparations, polypeptide/molecule affixed to a solid support, chemical libraries, or natural product mixtures. The assay may also simply comprise the steps of mixing a candidate compound with a solution containing a polypeptide, measuring polypeptide/molecule activity or binding, and comparing the polypeptide/molecule activity or binding to a standard.

Preferably, an ELISA assay can measure polypeptide level or activity in a sample (e.g., biological sample) using a monoclonal or polyclonal antibody. The antibody can measure polypeptide level or activity by either binding, directly or indirectly, to the polypeptide or by competing with the polypeptide for a substrate.

Additionally, the receptor to which the polypeptide of the present invention binds can be identified by numerous methods known to those of skill in the art, for example, ligand panning and FACS sorting (Coligan, et al., Current Protocols in Immun., 1(2), Chapter 5, (1991)). For example, expression cloning is employed wherein polyadenylated RNA is prepared from a cell responsive to the polypeptides, for example, NIH3T3 cells which are known to contain multiple receptors for the FGF family proteins, and SC-3 cells, and a cDNA library created from this RNA is divided into pools and used to transfect COS cells or other cells that are not responsive to the polypeptides. Transfected cells which are grown on glass slides are exposed to the polypeptide of the present invention, after they have been labeled. The polypeptides can be labeled by a variety of means including iodination or inclusion of a recognition site for a site-specific protein kinase.

Following fixation and incubation, the slides are subjected to auto-radiographic analysis. Positive pools are identified and sub-pools are prepared and re-transfected using an iterative sub-pooling and re-screening process, eventually yielding a single clones that encodes the putative receptor.

As an alternative approach for receptor identification, the labeled polypeptides can be photoaffinity linked with cell membrane or extract preparations that express the receptor molecule. Cross-linked material is resolved by PAGE analysis and exposed to X-ray film. The labeled complex containing the receptors of the polypeptides can be excised, resolved into peptide fragments, and subjected to protein microsequencing. The amino acid sequence obtained from microsequencing would be used to design a set of degenerate oligonucleotide probes to screen a cDNA library to identify the genes encoding the putative receptors.

Moreover, the techniques of gene-shuffling, motif-shuffling, exon-shuffling, and/or codon-shuffling (collectively referred to as “DNA shuffling”) may be employed to modulate the activities of the polypeptide of the present invention thereby effectively generating agonists and antagonists of the polypeptide of the present invention. See generally, U.S. Pat. Nos. 5,605,793, 5,811,238, 5,830,721, 5,834,252, and 5,837,458, and Patten, P. A., et al., Curr. Opinion Biotechnol. 8:724-33 (1997); Harayama, S. Trends Biotechnol. 16(2):76-82 (1998); Hansson, L. O., et al., J. Mol. Biol. 287:265-76 (1999); and Lorenzo, M. M. and Blasco, R. Biotechniques 24(2):308-13 (1998); each of these patents and publications are hereby incorporated by reference). In one embodiment, alteration of polynucleotides and corresponding polypeptides may be achieved by DNA shuffling. DNA shuffling involves the assembly of two or more DNA segments into a desired molecule by homologous, or site-specific, recombination. In another embodiment, polynucleotides and corresponding polypeptides may be altered by being subjected to random mutagenesis by error-prone PCR, random nucleotide insertion or other methods prior to recombination. In another embodiment, one or more components, motifs, sections, parts, domains, fragments, etc., of the polypeptide of the present invention may be recombined with one or more components, motifs, sections, parts, domains, fragments, etc. of one or more heterologous molecules. In preferred embodiments, the heterologous molecules are family members. In further preferred embodiments, the heterologous molecule is a growth factor such as, for example, platelet-derived growth factor (PDGF), insulin-like growth factor (IGF-I), transforming growth factor (TGF)-alpha, epidermal growth factor (EGF), fibroblast growth factor (FGF), TGF-beta, bone morphogenetic protein (BMP)-2, BMP-4, BMP-5, BMP-6, BMP-7, activins A and B, decapentaplegic (dpp), 60A, OP-2, dorsalin, growth differentiation factors (GDFs), nodal, MIS, inhibin-alpha, TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta5, and glial-derived neurotrophic factor (GDNF).

Other preferred fragments are biologically active fragments of the polypeptide of the present invention. Biologically active fragments are those exhibiting activity similar, but not necessarily identical, to an activity of the polypeptide of the present invention. The biological activity of the fragments may include an improved desired activity, or a decreased undesirable activity.

Additionally, this invention provides a method of screening compounds to identify those which modulate the action of the polypeptide of the present invention. An example of such an assay comprises combining a mammalian fibroblast cell, a the polypeptide of the present invention, the compound to be screened and ³[H] thymidine under cell culture conditions where the fibroblast cell would normally proliferate. A control assay may be performed in the absence of the compound to be screened and compared to the amount of fibroblast proliferation in the presence of the compound to determine if the compound stimulates proliferation by determining the uptake of ³[H] thymidine in each case. The amount of fibroblast cell proliferation is measured by liquid scintillation chromatography which measures the incorporation of ³[H] thymidine. Both agonist and antagonist compounds may be identified by this procedure.

In another method, a mammalian cell or membrane preparation expressing a receptor for a polypeptide of the present invention is incubated with a labeled polypeptide of the present invention in the presence of the compound. The ability of the compound to enhance or block this interaction could then be measured. Alternatively, the response of a known second messenger system following interaction of a compound to be screened and the receptor is measured and the ability of the compound to bind to the receptor and elicit a second messenger response is measured to determine if the compound is a potential agonist or antagonist. Such second messenger systems include but are not limited to, cAMP guanylate cyclase, ion channels or phosphoinositide hydrolysis.

All of these above assays can be used as diagnostic or prognostic markers. The molecules discovered using these assays can be used to treat disease or to bring about a particular result in a patient (e.g., blood vessel growth) by activating or inhibiting the polypeptide/molecule. Moreover, the assays can discover agents which may inhibit or enhance the production of the polypeptides of the invention from suitably manipulated cells or tissues.

Therefore, the invention includes a method of identifying compounds which bind to a polypeptide of the invention comprising the steps of: (a) incubating a candidate binding compound with a polypeptide of the present invention; and (b) determining if binding has occurred. Moreover, the invention includes a method of identifying agonists/antagonists comprising the steps of: (a) incubating a candidate compound with a polypeptide of the present invention, (b) assaying a biological activity, and (b) determining if a biological activity of the polypeptide has been altered.

Targeted Delivery

In another embodiment, the invention provides a method of delivering compositions to targeted cells expressing a receptor for a polypeptide of the invention, or cells expressing a cell bound form of a polypeptide of the invention.

As discussed herein, polypeptides or antibodies of the invention may be associated with heterologous polypeptides, heterologous nucleic acids, toxins, or prodrugs via hydrophobic, hydrophilic, ionic and/or covalent interactions. In one embodiment, the invention provides a method for the specific delivery of compositions of the invention to cells by administering polypeptides of the invention (including antibodies) that are associated with heterologous polypeptides or nucleic acids. In one example, the invention provides a method for delivering a therapeutic protein into the targeted cell. In another example, the invention provides a method for delivering a single stranded nucleic acid (e.g., antisense or ribozymes) or double stranded nucleic acid (e.g., DNA that can integrate into the cell's genome or replicate episomally and that can be transcribed) into the targeted cell.

In another embodiment, the invention provides a method for the specific destruction of cells (e.g., the destruction of tumor cells) by administering polypeptides of the invention (e.g., polypeptides of the invention or antibodies of the invention) in association with toxins or cytotoxic prodrugs.

By “toxin” is meant compounds that bind and activate endogenous cytotoxic effector systems, radioisotopes, holotoxins, modified toxins, catalytic subunits of toxins, or any molecules or enzymes not normally present in or on the surface of a cell that under defined conditions cause the cell's death. Toxins that may be used according to the methods of the invention include, but are not limited to, radioisotopes known in the art, compounds such as, for example, antibodies (or complement fixing containing portions thereof) that bind an inherent or induced endogenous cytotoxic effector system, thymidine kinase, endonuclease, RNAse, alpha toxin, ricin, abrin, Pseudomonas exotoxin A, diphtheria toxin, saporin, momordin, gelonin, pokeweed antiviral protein, alpha-sarcin and cholera toxin. By “cytotoxic prodrug” is meant a non-toxic compound that is converted by an enzyme, normally present in the cell, into a cytotoxic compound. Cytotoxic prodrugs that may be used according to the methods of the invention include, but are not limited to, glutamyl derivatives of benzoic acid mustard alkylating agent, phosphate derivatives of etoposide or mitomycin C, cytosine arabinoside, daunorubisin, and phenoxyacetamide derivatives of doxorubicin.

Drug Screening

Further contemplated is the use of the polypeptides of the present invention, or the polynucleotides encoding these polypeptides, to screen for molecules which modify the activities of the polypeptides of the present invention. Such a method would include contacting the polypeptide of the present invention with a selected compound(s) suspected of having antagonist or agonist activity, and assaying the activity of these polypeptides following binding.

This invention is particularly useful for screening therapeutic compounds by using the polypeptides of the present invention, or binding fragments thereof, in any of a variety of drug screening techniques. The polypeptide or fragment employed in such a test may be affixed to a solid support, expressed on a cell surface, free in solution, or located intracellularly. One method of drug screening utilizes eukaryotic or prokaryotic host cells which are stably transformed with recombinant nucleic acids expressing the polypeptide or fragment. Drugs are screened against such transformed cells in competitive binding assays. One may measure, for example, the formulation of complexes between the agent being tested and a polypeptide of the present invention.

Thus, the present invention provides methods of screening for drugs or any other agents which affect activities mediated by the polypeptides of the present invention. These methods comprise contacting such an agent with a polypeptide of the present invention or a fragment thereof and assaying for the presence of a complex between the agent and the polypeptide or a fragment thereof, by methods well known in the art. In such a competitive binding assay, the agents to screen are typically labeled. Following incubation, free agent is separated from that present in bound form, and the amount of free or uncomplexed label is a measure of the ability of a particular agent to bind to the polypeptides of the present invention.

Another technique for drug screening provides high throughput screening for compounds having suitable binding affinity to the polypeptides of the present invention, and is described in great detail in European Patent Application 84/03564, published on Sep. 13, 1984, which is incorporated herein by reference herein. Briefly stated, large numbers of different small peptide test compounds are synthesized on a solid substrate, such as plastic pins or some other surface. The peptide test compounds are reacted with polypeptides of the present invention and washed. Bound polypeptides are then detected by methods well known in the art. Purified polypeptides are coated directly onto plates for use in the aforementioned drug screening techniques. In addition, non-neutralizing antibodies may be used to capture the peptide and immobilize it on the solid support.

This invention also contemplates the use of competitive drug screening assays in which neutralizing antibodies capable of binding polypeptides of the present invention specifically compete with a test compound for binding to the polypeptides or fragments thereof. In this manner, the antibodies are used to detect the presence of any peptide which shares one or more antigenic epitopes with a polypeptide of the invention.

Antisense and Ribozyme (Antagonists)

In specific embodiments, antagonists according to the present invention are nucleic acids corresponding to the sequences contained in SEQ ID NO:X, or the complementary strand thereof, and/or to cDNA sequences contained in cDNA ATCC Deposit No:Z identified for example, in Table 1A and/or 1B. In one embodiment, antisense sequence is generated internally, by the organism, in another embodiment, the antisense sequence is separately administered (see, for example, O'Connor, J., Neurochem. 56:560 (1991). Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, Fla. (1988). Antisense technology can be used to control gene expression through antisense DNA or RNA, or through triple-helix formation. Antisense techniques are discussed for example, in Okano, J., Neurochem. 56:560 (1991); Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, Fla. (1988). Triple helix formation is discussed in, for instance, Lee et al., Nucleic Acids Research 6:3073 (1979); Cooney et al., Science 241:456 (1988); and Dervan et al., Science 251:1300 (1991). The methods are based on binding of a polynucleotide to a complementary DNA or RNA.

For example, the use of c-myc and c-myb antisense RNA constructs to inhibit the growth of the non-lymphocytic leukemia cell line HL-60 and other cell lines was previously described. (Wickstrom et al. (1988); Anfossi et al. (1989)). These experiments were performed in vitro by incubating cells with the oligoribonucleotide. A similar procedure for in vivo use is described in WO 91/15580. Briefly, a pair of oligonucleotides for a given antisense RNA is produced as follows: A sequence complimentary to the first 15 bases of the open reading frame is flanked by an EcoR1 site on the 5 end and a HindIII site on the 3 end. Next, the pair of oligonucleotides is heated at 90° C. for one minute and then annealed in 2× ligation buffer (20 mM TRIS HCl pH 7.5, 10 mM MgCl2, 10 MM dithiothreitol (DTT) and 0.2 mM ATP) and then ligated to the EcoR1/Hind III site of the retroviral vector PMV7 (WO 91/15580).

For example, the 5′ coding portion of a polynucleotide that encodes the polypeptide of the present invention may be used to design an antisense RNA oligonucleotide of from about 10 to 40 base pairs in length. A DNA oligonucleotide is designed to be complementary to a region of the gene involved in transcription thereby preventing transcription and the production of the receptor. The antisense RNA oligonucleotide hybridizes to the mRNA in vivo and blocks translation of the mRNA molecule into receptor polypeptide.

In one embodiment, the antisense nucleic acid of the invention is produced intracellularly by transcription from an exogenous sequence. For example, a vector or a portion thereof, is transcribed, producing an antisense nucleic acid (RNA) of the invention. Such a vector would contain a sequence encoding the antisense nucleic acid. Such a vector can remain episomal or become chromosomally integrated, as long as it can be transcribed to produce the desired antisense RNA. Such vectors can be constructed by recombinant DNA technology methods standard in the art. Vectors can be plasmid, viral, or others known in the art, used for replication and expression in vertebrate cells. Expression of the sequence encoding the polypeptide of the present invention or fragments thereof, can be by any promoter known in the art to act in vertebrate, preferably human cells. Such promoters can be inducible or constitutive. Such promoters include, but are not limited to, the SV40 early promoter region (Bernoist and Chambon, Nature 29:304-310 (1981), the promoter contained in the 3′ long terminal repeat of Rous sarcoma virus (Yamamoto et al., Cell 22:787-797 (1980), the herpes thymidine promoter (Wagner et al., Proc. Natl. Acad. Sci. U.S.A. 78:1441-1445 (1981), the regulatory sequences of the metallothionein gene (Brinster, et al., Nature 296:39-42 (1982)), etc.

The antisense nucleic acids of the invention comprise a sequence complementary to at least a portion of an RNA transcript of a gene of the present invention. However, absolute complementarity, although preferred, is not required. A sequence “complementary to at least a portion of an RNA,” referred to herein, means a sequence having sufficient complementarity to be able to hybridize with the RNA, forming a stable duplex; in the case of double stranded antisense nucleic acids, a single strand of the duplex DNA may thus be tested, or triplex formation may be assayed. The ability to hybridize will depend on both the degree of complementarity and the length of the antisense nucleic acid. Generally, the larger the hybridizing nucleic acid, the more base mismatches with a RNA it may contain and still form a stable duplex (or triplex as the case may be). One skilled in the art can ascertain a tolerable degree of mismatch by use of standard procedures to determine the melting point of the hybridized complex.

Oligonucleotides that are complementary to the 5′ end of the message, e.g., the 5′ untranslated sequence up to and including the AUG initiation codon, should work most efficiently at inhibiting translation. However, sequences complementary to the 3′ untranslated sequences of mRNAs have been shown to be effective at inhibiting translation of mRNAs as well. See generally, Wagner, R., 1994, Nature 372:333-335. Thus, oligonucleotides complementary to either the 5′- or 3′-non-translated, non-coding regions of polynucleotide sequences described herein could be used in an antisense approach to inhibit translation of endogenous mRNA. Oligonucleotides complementary to the 5′ untranslated region of the mRNA should include the complement of the AUG start codon. Antisense oligonucleotides complementary to mRNA coding regions are less efficient inhibitors of translation but could be used in accordance with the invention. Whether designed to hybridize to the 5′-, 3′- or coding region of mRNA of the present invention, antisense nucleic acids should be at least six nucleotides in length, and are preferably oligonucleotides ranging from 6 to about 50 nucleotides in length. In specific aspects the oligonucleotide is at least 10 nucleotides, at least 17 nucleotides, at least 25 nucleotides or at least 50 nucleotides.

The polynucleotides of the invention can be DNA or RNA or chimeric mixtures or derivatives or modified versions thereof, single-stranded or double-stranded. The oligonucleotide can be modified at the base moiety, sugar moiety, or phosphate backbone, for example, to improve stability of the molecule, hybridization, etc. The oligonucleotide may include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger et al., 1989, Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556; Lemaitre et al., 1987, Proc. Natl. Acad. Sci. 84:648-652; PCT Publication No. WO88/09810, published Dec. 15, 1988) or the blood-brain barrier (see, e.g., PCT Publication No. WO89/10134, published Apr. 25, 1988), hybridization-triggered cleavage agents. (See, e.g., Krol et al., 1988, BioTechniques 6:958-976) or intercalating agents. (See, e.g., Zon, 1988, Pharm. Res. 5:539-549). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

The antisense oligonucleotide may comprise at least one modified base moiety which is selected from the group including, but not limited to, 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xantine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5′-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N-6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine.

The antisense oligonucleotide may also comprise at least one modified sugar moiety selected from the group including, but not limited to, arabinose, 2-fluoroarabinose, xylulose, and hexose.

In yet another embodiment, the antisense oligonucleotide comprises at least one modified phosphate backbone selected from the group including, but not limited to, a phosphorothioate, a phosphorodithioate, a phosphoramidothioate, a phosphoramidate, a phosphordiamidate, a methylphosphonate, an alkyl phosphotriester, and a formacetal or analog thereof.

In yet another embodiment, the antisense oligonucleotide is an a-anomeric oligonucleotide. An a-anomeric oligonucleotide forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual b-units, the strands run parallel to each other (Gautier et al., 1987, Nucl. Acids Res. 15:6625-6641). The oligonucleotide is a 2′-0-methylribonucleotide (Inoue et al., 1987, Nucl. Acids Res. 15:6131-6148), or a chimeric RNA-DNA analogue (Inoue et al., 1987, FEBS Lett. 215:327-330).

Polynucleotides of the invention may be synthesized by standard methods known in the art, e.g. by use of an automated DNA synthesizer (such as are commercially available from Biosearch, Applied Biosystems, etc.). As examples, phosphorothioate oligonucleotides may be synthesized by the method of Stein et al. (1988, Nucl. Acids Res. 16:3209), methylphosphonate oligonucleotides can be prepared by use of controlled pore glass polymer supports (Sarin et al., 1988, Proc. Natl. Acad. Sci. U.S.A. 85:7448-7451), etc.

While antisense nucleotides complementary to the coding region sequence could be used, those complementary to the transcribed untranslated region are most preferred.

Potential antagonists according to the invention also include catalytic RNA, or a ribozyme (Sce, e.g., PCT International Publication WO 90/11364, published Oct. 4, 1990; Sarver et al, Science 247:1222-1225 (1990). While ribozymes that cleave mRNA at site specific recognition sequences can be used to destroy mRNAs, the use of hammerhead ribozymes is preferred. Hammerhead ribozymes cleave mRNAs at locations dictated by flanking regions that form complementary base pairs with the target mRNA. The sole requirement is that the target mRNA have the following sequence of two bases: 5′-UG-3′. The construction and production of hammerhead ribozymes is well known in the art and is described more fully in Haseloff and Gerlach, Nature 334:585-591 (1988). There are numerous potential hammerhead ribozyme cleavage sites within the nucleotide sequence of SEQ ID NO:X. Preferably, the ribozyme is engineered so that the cleavage recognition site is located near the 5′ end of the mRNA; i.e., to increase efficiency and minimize the intracellular accumulation of non-functional mRNA transcripts.

As in the antisense approach, the ribozymes of the invention can be composed of modified oligonucleotides (e.g., for improved stability, targeting, etc.) and should be delivered to cells which express in vivo. DNA constructs encoding the ribozyme may be introduced into the cell in the same manner as described above for the introduction of antisense encoding DNA. A preferred method of delivery involves using a DNA construct “encoding” the ribozyme under the control of a strong constitutive promoter, such as, for example, pol III or pol II promoter, so that transfected cells will produce sufficient quantities of the ribozyme to destroy endogenous messages and inhibit translation. Since ribozymes unlike antisense molecules, are catalytic, a lower intracellular concentration is required for efficiency.

Antagonist/agonist compounds may be employed to inhibit the cell growth and proliferation effects of the polypeptides of the present invention on neoplastic cells and tissues, i.e. stimulation of angiogenesis of tumors, and, therefore, retard or prevent abnormal cellular growth and proliferation, for example, in tumor formation or growth.

The antagonist/agonist may also be employed to prevent hyper-vascular diseases, and prevent the proliferation of epithelial lens cells after extracapsular cataract surgery. Prevention of the mitogenic activity of the polypeptides of the present invention may also be desirous in cases such as restenosis after balloon angioplasty.

The antagonist/agonist may also be employed to prevent the growth of scar tissue during wound healing.

The antagonist/agonist may also be employed to treat the diseases described herein.

Thus, the invention provides a method of treating disorders or diseases, including but not limited to the disorders or diseases listed throughout this application, associated with overexpression of a polynucleotide of the present invention by administering to a patient (a) an antisense molecule directed to the polynucleotide of the present invention, and/or (b) a ribozyme directed to the polynucleotide of the present invention.

Binding Peptides and Other Molecules

The invention also encompasses screening methods for identifying polypeptides and nonpolypeptides that bind polypeptides of the invention, and the binding molecules identified thereby. These binding molecules are useful, for example, as agonists and antagonists of the polypeptides of the invention. Such agonists and antagonists can be used, in accordance with the invention, in the therapeutic embodiments described in detail, below.

This method comprises the steps of:

-   -   a. contacting polypeptides of the invention with a plurality of         molecules; and     -   b. identifying a molecule that binds the polypeptides of the         invention.

The step of contacting the polypeptides of the invention with the plurality of molecules may be effected in a number of ways. For example, one may contemplate immobilizing the polypeptides on a solid support and bringing a solution of the plurality of molecules in contact with the immobilized polypeptides. Such a procedure would be akin to an affinity chromatographic process, with the affinity matrix being comprised of the immobilized polypeptides of the invention. The molecules having a selective affinity for the polypeptides can then be purified by affinity selection. The nature of the solid support, process for attachment of the polypeptides to the solid support, solvent, and conditions of the affinity isolation or selection are largely conventional and well known to those of ordinary skill in the art.

Alternatively, one may also separate a plurality of polypeptides into substantially separate fractions comprising a subset of or individual polypeptides. For instance, one can separate the plurality of polypeptides by gel electrophoresis, column chromatography, or like method known to those of ordinary skill for the separation of polypeptides. The individual polypeptides can also be produced by a transformed host cell in such a way as to be expressed on or about its outer surface (e.g., a recombinant phage). Individual isolates can then be “probed” by the polypeptides of the invention, optionally in the presence of an inducer should one be required for expression, to determine if any selective affinity interaction takes place between the polypeptides and the individual clone. Prior to contacting the polypeptides with each fraction comprising individual polypeptides, the polypeptides could first be transferred to a solid support for additional convenience. Such a solid support may simply be a piece of filter membrane, such as one made of nitrocellulose or nylon. In this manner, positive clones could be identified from a collection of transformed host cells of an expression library, which harbor a DNA construct encoding a polypeptide having a selective affinity for polypeptides of the invention. Furthermore, the amino acid sequence of the polypeptide having a selective affinity for the polypeptides of the invention can be determined directly by conventional means or the coding sequence of the DNA encoding the polypeptide can frequently be determined more conveniently. The primary sequence can then be deduced from the corresponding DNA sequence. If the amino acid sequence is to be determined from the polypeptide itself, one may use microsequencing techniques. The sequencing technique may include mass spectroscopy.

In certain situations, it may be desirable to wash away any unbound polypeptides from a mixture of the polypeptides of the invention and the plurality of polypeptides prior to attempting to determine or to detect the presence of a selective affinity interaction. Such a wash step may be particularly desirable when the polypeptides of the invention or the plurality of polypeptides are bound to a solid support.

The plurality of molecules provided according to this method may be provided by way of diversity libraries, such as random or combinatorial peptide or nonpeptide libraries which can be screened for molecules that specifically bind polypeptides of the invention. Many libraries are known in the art that can be used, e.g., chemically synthesized libraries, recombinant (e.g., phage display libraries), and in vitro translation-based libraries. Examples of chemically synthesized libraries are described in Fodor et al., 1991, Science 251:767-773; Houghten et al., 1991, Nature 354:84-86; Lam et al., 1991, Nature 354:82-84; Medynski, 1994, Bio/Technology 12:709-710; Gallop et al., 1994, J. Medicinal Chemistry 37(9):1233-1251; Ohlmeyer et al., 1993, Proc. Natl. Acad. Sci. USA 90:10922-10926; Erb et al., 1994, Proc. Natl. Acad. Sci. USA 91:11422-11426; Houghten et al., 1992, Biotechniques 13:412; Jayawickreme et al., 1994, Proc. Natl. Acad. Sci. USA 91:1614-1618; Salmon et al., 1993, Proc. Natl. Acad. Sci. USA 90:11708-11712; PCT Publication No. WO 93/20242; and Brenner and Lerner, 1992, Proc. Natl. Acad. Sci. USA 89:5381-5383.

Examples of phage display libraries are described in Scott and Smith, 1990, Science 249:386-390; Devlin et al., 1990, Science, 249:404-406; Christian, R. B., et al., 1992, J. Mol. Biol. 227:711-718); Lenstra, 1992, J. Immunol. Meth. 152:149-157; Kay et al., 1993, Gene 128:59-65; and PCT Publication No. WO 94/18318 dated Aug. 18, 1994.

In vitro translation-based libraries include but are not limited to those described in PCT Publication No. WO 91/05058 dated Apr. 18, 1991; and Mattheakis et al., 1994, Proc. Natl. Acad. Sci. USA 91:9022-9026.

By way of examples of nonpeptide libraries, a benzodiazepine library (see e.g., Bunin et al., 1994, Proc. Natl. Acad. Sci. USA 91:4708-4712) can be adapted for use. Peptoid libraries (Simon et al., 1992, Proc. Natl. Acad. Sci. USA 89:9367-9371) can also be used. Another example of a library that can be used, in which the amide functionalities in peptides have been permethylated to generate a chemically transformed combinatorial library, is described by Ostresh et al. (1994, Proc. Natl. Acad. Sci. USA 91:11138-11142).

The variety of non-peptide libraries that are useful in the present invention is great. For example, Ecker and Crooke, 1995, Bio/Technology 13:351-360 list benzodiazepines, hydantoins, piperazinediones, biphenyls, sugar analogs, beta-mercaptoketones, arylacetic acids, acylpiperidines, benzopyrans, cubanes, xanthines, aminimides, and oxazolones as among the chemical species that form the basis of various libraries.

Non-peptide libraries can be classified broadly into two types: decorated monomers and oligomers. Decorated monomer libraries employ a relatively simple scaffold structure upon which a variety functional groups is added. Often the scaffold will be a molecule with a known useful pharmacological activity. For example, the scaffold might be the benzodiazepine structure.

Non-peptide oligomer libraries utilize a large number of monomers that are assembled together in ways that create new shapes that depend on the order of the monomers. Among the monomer units that have been used are carbamates, pyrrolinones, and morpholinos. Peptoids, peptide-like oligomers in which the side chain is attached to the alpha amino group rather than the alpha carbon, form the basis of another version of non-peptide oligomer libraries. The first non-peptide oligomer libraries utilized a single type of monomer and thus contained a repeating backbone. Recent libraries have utilized more than one monomer, giving the libraries added flexibility.

Screening the libraries can be accomplished by any of a variety of commonly known methods. See, e.g., the following references, which disclose screening of peptide libraries: Parmley and Smith, 1989, Adv. Exp. Med. Biol. 251:215-218; Scott and Smith, 1990, Science 249:386-390; Fowlkes et al., 1992; BioTechniques 13:422-427; Oldenburg et al., 1992, Proc. Natl. Acad. Sci. USA 89:5393-5397; Yu et al., 1994, Cell 76:933-945; Staudt et al., 1988, Science 241:577-580; Bock et al., 1992, Nature 355:564-566; Tuerk et al., 1992, Proc. Natl. Acad. Sci. USA 89:6988-6992; Ellington et al., 1992, Nature 355:850-852; U.S. Pat. No. 5,096,815, U.S. Pat. No. 5,223,409, and U.S. Pat. No. 5,198,346, all to Ladner et al.; Rebar and Pabo, 1993, Science 263:671-673; and CT Publication No. WO 94/18318.

In a specific embodiment, screening to identify a molecule that binds polypeptides of the invention can be carried out by contacting the library members with polypeptides of the invention immobilized on a solid phase and harvesting those library members that bind to the polypeptides of the invention. Examples of such screening methods, termed “panning” techniques are described by way of example in Parmley and Smith, 1988, Gene 73:305-318; Fowlkes et al., 1992, BioTechniques 13:422-427; PCT Publication No. WO 94/18318; and in references cited herein.

In another embodiment, the two-hybrid system for selecting interacting proteins in yeast (Fields and Song, 1989, Nature 340:245-246; Chien et al., 1991, Proc. Natl. Acad. Sci. USA 88:9578-9582) can be used to identify molecules that specifically bind to polypeptides of the invention.

Where the binding molecule is a polypeptide, the polypeptide can be conveniently selected from any peptide library, including random peptide libraries, combinatorial peptide libraries, or biased peptide libraries. The term “biased” is used herein to mean that the method of generating the library is manipulated so as to restrict one or more parameters that govern the diversity of the resulting collection of molecules, in this case peptides.

Thus, a truly random peptide library would generate a collection of peptides in which the probability of finding a particular amino acid at a given position of the peptide is the same for all 20 amino acids. A bias can be introduced into the library, however, by specifying, for example, that a lysine occur every fifth amino acid or that positions 4, 8, and 9 of a decapeptide library be fixed to include only arginine. Clearly, many types of biases can be contemplated, and the present invention is not restricted to any particular bias. Furthermore, the present invention contemplates specific types of peptide libraries, such as phage displayed peptide libraries and those that utilize a DNA construct comprising a lambda phage vector with a DNA insert.

As mentioned above, in the case of a binding molecule that is a polypeptide, the polypeptide may have about 6 to less than about 60 amino acid residues, preferably about 6 to about 10 amino acid residues, and most preferably, about 6 to about 22 amino acids. In another embodiment, a binding polypeptide has in the range of 15-100 amino acids, or 20-50 amino acids.

The selected binding polypeptide can be obtained by chemical synthesis or recombinant expression.

Other Activities

A polypeptide, polynucleotide, agonist, or antagonist of the present invention, as a result of the ability to stimulate vascular endothelial cell growth, may be employed in treatment for stimulating re-vascularization of ischemic tissues due to various disease conditions such as thrombosis, arteriosclerosis, and other cardiovascular conditions. The polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be employed to stimulate angiogenesis and limb regeneration, as discussed above.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be employed for treating wounds due to injuries, burns, post-operative tissue repair, and ulcers since they are mitogenic to various cells of different origins, such as fibroblast cells and skeletal muscle cells, and therefore, facilitate the repair or replacement of damaged or diseased tissue.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be employed stimulate neuronal growth and to treat and prevent neuronal damage which occurs in certain neuronal disorders or neuro-degenerative conditions such as Alzheimer's disease, Parkinson's disease, and AIDS-related complex. A polypeptide, polynucleotide, agonist, or antagonist of the present invention may have the ability to stimulate chondrocyte growth, therefore, they may be employed to enhance bone and periodontal regeneration and aid in tissue transplants or bone grafts.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may be also be employed to prevent skin aging due to sunburn by stimulating keratinocyte growth.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be employed for preventing hair loss, since FGF family members activate hair-forming cells and promotes melanocyte growth. Along the same lines, a polypeptide, polynucleotide, agonist, or antagonist of the present invention may be employed to stimulate growth and differentiation of hematopoietic cells and bone marrow cells when used in combination with other cytokines.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be employed to maintain organs before transplantation or for supporting cell culture of primary tissues. A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be employed for inducing tissue of mesodermal origin to differentiate in early embryos.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also increase or decrease the differentiation or proliferation of embryonic stem cells, besides, as discussed above, hematopoietic lineage.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be used to modulate mammalian characteristics, such as body height, weight, hair color, eye color, skin, percentage of adipose tissue, pigmentation, size, and shape (e.g., cosmetic surgery). Similarly, a polypeptide, polynucleotide, agonist, or antagonist of the present invention may be used to modulate mammalian metabolism affecting catabolism, anabolism, processing, utilization, and storage of energy.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may be used to change a mammal's mental state or physical state by influencing biorhythms, caricadic rhythms, depression (including depressive disorders), tendency for violence, tolerance for pain, reproductive capabilities (preferably by Activin or Inhibin-like activity), hormonal or endocrine levels, appetite, libido, memory, stress, or other cognitive qualities.

A polypeptide, polynucleotide, agonist, or antagonist of the present invention may also be used as a food additive or preservative, such as to increase or decrease storage capabilities, fat content, lipid, protein, carbohydrate, vitamins, minerals, cofactors or other nutritional components.

The above-recited applications have uses in a wide variety of hosts. Such hosts include, but are not limited to, human, murine, rabbit, goat, guinea pig, camel, horse, mouse, rat, hamster, pig, micro-pig, chicken, goat, cow, sheep, dog, cat, non-human primate, and human. In specific embodiments, the host is a mouse, rabbit, goat, guinea pig, chicken, rat, hamster, pig, sheep, dog or cat. In preferred embodiments, the host is a mammal. In most preferred embodiments, the host is a human.

Other Preferred Embodiments

Other preferred embodiments of the claimed invention include an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a sequence of at least about 50 contiguous nucleotides in the nucleotide sequence of SEQ ID NO:X or the complementary strand thereto, the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto, and/or cDNA contained in ATCC Deposit No:Z.

Also preferred is a nucleic acid molecule wherein said sequence of contiguous nucleotides is included in the nucleotide sequence of the portion of SEQ ID NO:X as defined in column 5, “ORF (From-To)”, in Table 1B.1.

Also preferred is a nucleic acid molecule wherein said sequence of contiguous nucleotides is included in the nucleotide sequence of the portion of SEQ ID NO:X as defined in columns 8 and 9, “NT From” and “NT To” respectively, in Table 2.

Also preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a sequence of at least about 150 contiguous nucleotides in the nucleotide sequence of SEQ ID NO:X or the complementary strand thereto, the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto, and/or cDNA contained in ATCC Deposit No:Z.

Further preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a sequence of at least about 500 contiguous nucleotides in the nucleotide sequence of SEQ ID NO:X or the complementary strand thereto, the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto, and/or cDNA contained in ATCC Deposit No:Z.

A further preferred embodiment is a nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the nucleotide sequence of the portion of SEQ ID NO:X defined in column 5, “ORF (From-To)”, in Table 1B.1.

A further preferred embodiment is a nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the nucleotide sequence of the portion of SEQ ID NO:X defined in columns 8 and 9, “NT From” and “NT To”, respectively, in Table 2.

A further preferred embodiment is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the complete nucleotide sequence of SEQ ID NO:X or the complementary strand thereto, the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto, and/or cDNA contained in ATCC Deposit No:Z.

Also preferred is an isolated nucleic acid molecule which hybridizes under stringent hybridization conditions to a nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:X or the complementary strand thereto, the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto, and/or cDNA contained in ATCC Deposit No:Z, wherein said nucleic acid molecule which hybridizes does not hybridize under stringent hybridization conditions to a nucleic acid molecule having a nucleotide sequence consisting of only A residues or of only T residues.

Also preferred is a composition of matter comprising a DNA molecule which comprises the cDNA contained in ATCC Deposit No:Z.

Also preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a sequence of at least 50 contiguous nucleotides of the cDNA sequence contained in ATCC Deposit No:Z.

Also preferred is an isolated nucleic acid molecule, wherein said sequence of at least 50 contiguous nucleotides is included in the nucleotide sequence of an open reading frame sequence encoded by cDNA contained in ATCC Deposit No:Z.

Also preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to sequence of at least 150 contiguous nucleotides in the nucleotide sequence encoded by cDNA contained in ATCC Deposit No:Z.

A further preferred embodiment is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to sequence of at least 500 contiguous nucleotides in the nucleotide sequence encoded by cDNA contained in ATCC Deposit No:Z.

A further preferred embodiment is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the complete nucleotide sequence encoded by cDNA contained in ATCC Deposit No:Z.

A further preferred embodiment is a method for detecting in a biological sample a nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of: a nucleotide sequence of SEQ ID NO:X or the complementary strand thereto; the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto; and a nucleotide sequence encoded by cDNA contained in ATCC Deposit No:Z; which method comprises a step of comparing a nucleotide sequence of at least one nucleic acid molecule in said sample with a sequence selected from said group and determining whether the sequence of said nucleic acid molecule in said sample is at least 95% identical to said selected sequence.

Also preferred is the above method wherein said step of comparing sequences comprises determining the extent of nucleic acid hybridization between nucleic acid molecules in said sample and a nucleic acid molecule comprising said sequence selected from said group. Similarly, also preferred is the above method wherein said step of comparing sequences is performed by comparing the nucleotide sequence determined from a nucleic acid molecule in said sample with said sequence selected from said group. The nucleic acid molecules can comprise DNA molecules or RNA molecules.

A further preferred embodiment is a method for identifying the species, tissue or cell type of a biological sample which method comprises a step of detecting nucleic acid molecules in said sample, if any, comprising a nucleotide sequence that is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of: a nucleotide sequence of SEQ ID NO:X or the complementary strand thereto; the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto; and a nucleotide sequence of the cDNA contained in ATCC Deposit No:Z.

The method for identifying the species, tissue or cell type of a biological sample can comprise a step of detecting nucleic acid molecules comprising a nucleotide sequence in a panel of at least two nucleotide sequences, wherein at least one sequence in said panel is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from said group.

Also preferred is a method for diagnosing in a subject a pathological condition associated with abnormal structure or expression of a nucleotide sequence of SEQ ID NO:X or the complementary strand thereto; the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto; or the cDNA contained in ATCC Deposit No:Z which encodes a protein, wherein the method comprises a step of detecting in a biological sample obtained from said subject nucleic acid molecules, if any, comprising a nucleotide sequence that is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of: a nucleotide sequence of SEQ ID NO:X or the complementary strand thereto; the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto; and a nucleotide sequence of cDNA contained in ATCC Deposit No:Z.

The method for diagnosing a pathological condition can comprise a step of detecting nucleic acid molecules comprising a nucleotide sequence in a panel of at least two nucleotide sequences, wherein at least one sequence in said panel is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from said group.

Also preferred is a composition of matter comprising isolated nucleic acid molecules wherein the nucleotide sequences of said nucleic acid molecules comprise a panel of at least two nucleotide sequences, wherein at least one sequence in said panel is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of: a nucleotide sequence of SEQ ID NO:X or the complementary strand thereto; the nucleotide sequence as defined in column 5 of Table 1B.1 or columns 8 and 9 of Table 2 or the complementary strand thereto; and a nucleotide sequence encoded by cDNA contained in ATCC Deposit No:Z. The nucleic acid molecules can comprise DNA molecules or RNA molecules.

Also preferred is a composition of matter comprising isolated nucleic acid molecules wherein the nucleotide sequences of said nucleic acid molecules comprise a DNA microarray or “chip” of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 100, 150, 200, 250, 300, 500, 1000, 2000, 3000, or 4000 nucleotide sequences, wherein at least one sequence in said DNA microarray or “chip” is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of: a nucleotide sequence of SEQ ID NO:X wherein X is any integer as defined in Table 1A and/or 1B; and a nucleotide sequence encoded by a human cDNA clone identified by a cDNA “Clone ID” in Table 1A and/or 1B.

Also preferred is an isolated polypeptide comprising an amino acid sequence at least 90% identical to a sequence of at least about 10 contiguous amino acids in the polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and/or a polypeptide encoded by cDNA contained in ATCC Deposit No:Z.

Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 30 contiguous amino acids in the amino acid sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and/or a polypeptide encoded by cDNA contained in ATCC Deposit No:Z.

Further preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 100 contiguous amino acids in the amino acid sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and/or a polypeptide encoded by cDNA contained in ATCC Deposit No:Z.

Further preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to the complete amino acid sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and/or a polypeptide encoded by cDNA contained in ATCC Deposit No:Z.

Further preferred is an isolated polypeptide comprising an amino acid sequence at least 90% identical to a sequence of at least about 10 contiguous amino acids in the complete amino acid sequence of a polypeptide encoded by contained in ATCC Deposit No:Z

Also preferred is a polypeptide wherein said sequence of contiguous amino acids is included in the amino acid sequence of a portion of said polypeptide encoded by cDNA contained in ATCC Deposit No:Z; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and/or the polypeptide sequence of SEQ ID NO:Y.

Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 30 contiguous amino acids in the amino acid sequence of a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 100 contiguous amino acids in the amino acid sequence of a polypeptide encoded by cDNA contained in ATCC Deposit No:Z.

Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to the amino acid sequence of a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

Further preferred is an isolated antibody which binds specifically to a polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: a polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

Further preferred is a method for detecting in a biological sample a polypeptide comprising an amino acid sequence which is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: a polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z; which method comprises a step of comparing an amino acid sequence of at least one polypeptide molecule in said sample with a sequence selected from said group and determining whether the sequence of said polypeptide molecule in said sample is at least 90% identical to said sequence of at least 10 contiguous amino acids.

Also preferred is the above method wherein said step of comparing an amino acid sequence of at least one polypeptide molecule in said sample with a sequence selected from said group comprises determining the extent of specific binding of polypeptides in said sample to an antibody which binds specifically to a polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: a polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

Also preferred is the above method wherein said step of comparing sequences is performed by comparing the amino acid sequence determined from a polypeptide molecule in said sample with said sequence selected from said group.

Also preferred is a method for identifying the species, tissue or cell type of a biological sample which method comprises a step of detecting polypeptide molecules in said sample, if any, comprising an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID. NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

Also preferred is the above method for identifying the species, tissue or cell type of a biological sample, which method comprises a step of detecting polypeptide molecules comprising an amino acid sequence in a panel of at least two amino acid sequences, wherein at least one sequence in said panel is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the above group.

Also preferred is a method for diagnosing in a subject a pathological condition associated with abnormal structure or expression of a nucleic acid sequence identified in Table 1A, 1B or Table 2 encoding a polypeptide, which method comprises a step of detecting in a biological sample obtained from said subject polypeptide molecules comprising an amino acid sequence in a panel of at least two amino acid sequences, wherein at least one sequence in said panel is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

In any of these methods, the step of detecting said polypeptide molecules includes using an antibody.

Also preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a nucleotide sequence encoding a polypeptide wherein said polypeptide comprises an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

Also preferred is an isolated nucleic acid molecule, wherein said nucleotide sequence encoding a polypeptide has been optimized for expression of said polypeptide in a prokaryotic host.

Also preferred is a polypeptide molecule, wherein said polypeptide comprises an amino acid sequence selected from the group consisting of: polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z.

Further preferred is a method of making a recombinant vector comprising inserting any of the above isolated nucleic acid molecule into a vector. Also preferred is the recombinant vector produced by this method. Also preferred is a method of making a recombinant host cell comprising introducing the vector into a host cell, as well as the recombinant host cell produced by this method.

Also preferred is a method of making an isolated polypeptide comprising culturing this recombinant host cell under conditions such that said polypeptide is expressed and recovering said polypeptide. Also preferred is this method of making an isolated polypeptide, wherein said recombinant host cell is a eukaryotic cell and said polypeptide is a human protein comprising an amino acid sequence selected from the group consisting of: polypeptide sequence of SEQ ID NO:Y; a polypeptide encoded by SEQ ID NO:X or the complementary strand thereto; the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2; and a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z. The isolated polypeptide produced by this method is also preferred.

Also preferred is a method of treatment of an individual in need of an increased level of a protein activity, which method comprises administering to such an individual a Therapeutic comprising an amount of an isolated polypeptide, polynucleotide, immunogenic fragment or analogue thereof, binding agent, antibody, or antigen binding fragment of the claimed invention effective to increase the level of said protein activity in said individual.

Also preferred is a method of treatment of an individual in need of a decreased level of a protein activity, which method comprised administering to such an individual a Therapeutic comprising an amount of an isolated polypeptide, polynucleotide, immunogenic fragment or analogue thereof, binding agent, antibody, or antigen binding fragment of the claimed invention effective to decrease the level of said protein activity in said individual.

Also preferred is a method of treatment of an individual in need of a specific delivery of toxic compositions to diseased cells (e.g., tumors, leukemias or lymphomas), which method comprises administering to such an individual a Therapeutic comprising an amount of an isolated polypeptide of the invention, including, but not limited to a binding agent, or antibody of the claimed invention that are associated with toxin or cytotoxic prodrugs.

Having generally described the invention, the same will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended as limiting.

Description of Table 6

Table 6 summarizes some of the ATCC Deposits, Deposit dates, and ATCC designation numbers of deposits made with the ATCC in connection with the present application. These deposits were made in addition to those described in the Table 1A. TABLE 6 ATCC Deposits Deposit Date ATCC Designation Number LP01, LP02, LP03, LP04, May-20-97 209059, 209060, 209061, LP05, LP06, LP07, LP08, 209062, 209063, 209064, LP09, LP10, LP11, 209065, 209066, 209067, 209068, 209069 LP12 Jan-12-98 209579 LP13 Jan-12-98 209578 LP14 Jul-16-98 203067 LP15 Jul-16-98 203068 LP16 Feb-1-99 203609 LP17 Feb-1-99 203610 LP20 Nov-17-98 203485 LP21 Jun-18-99 PTA-252 LP22 Jun-18-99 PTA-253 LP23 Dec-22-99 PTA-1081

EXAMPLES Example 1 Isolation of a Selected cDNA Clone Front the Deposited Sample

Each ATCC Deposit No:Z is contained in a plasmid vector. Table 7 identifies the vectors used to construct the cDNA library from which each clone was isolated. In many cases, the vector used to construct the library is a phage vector from which a plasmid has been excised. The following correlates the related plasmid for each phage vector used in constructing the cDNA library. For example, where a particular clone is identified in Table 7 as being isolated in the vector “Lambda Zap,” the corresponding deposited clone is in “pBluescript.” Vector Used to Construct Library Corresponding Deposited Plasmid Lambda Zap pBluescript (pBS) Uni-Zap XR pBluescript (pBS) Zap Express pBK lafmid BA plafmid BA pSport1 pSport1 pCMVSport 2.0 pCMVSport 2.0 pCMVSport 3.0 pCMVSport 3.0 pCR ® 2.1 pCR ® 2.1

Vectors Lambda Zap (U.S. Pat. Nos. 5,128,256 and 5,286,636), Uni-Zap XR (U.S. Pat. Nos. 5,128,256 and 5,286,636), Zap Express (U.S. Pat. Nos. 5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. et al., Nucleic Acids Res. 16:7583-7600 (1988); Alting-Mees, M. A. and Short, J. M., Nucleic Acids Res. 17:9494 (1989)) and pBK (Alting-Mees, M. A. et al., Strategies 5:58-61 (1992)) are commercially available from Stratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La Jolla, Calif., 92037. pBS contains an ampicillin resistance gene and pBK contains a neomycin resistance gene. Both can be transformed into E. coli strain XL-1 Blue, also available from Stratagene. pBS comes in 4 forms SK+, SK−, KS+ and KS. The S and K refers to the orientation of the polylinker to the T7 and T3 primer sequences which flank the polylinker region (“S” is for SacI and “K” is for KpnI which are the first sites on each respective end of the linker). “+” or “−” refer to the orientation of the f1 origin of replication (“ori”), such that in one orientation, single stranded rescue initiated from the f1 ori generates sense strand DNA and in the other, antisense.

Vectors pSport1, pCMVSport 2.0 and pCMVSport 3.0, were obtained from Life Technologies, Inc., P.O. Box 6009, Gaithersburg, Md. 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. (See, for instance, Gruber, C. E., et al., Focus 15:59 (1993)). Vector lafmid BA (Bento Soares, Columbia University, NY) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue. Vector pCR®2.1, which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, Calif. 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. (See, for instance, Clark, J. M., Nuc. Acids Res. 16:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991)). Preferably, a polynucleotide of the present invention does not comprise the phage vector sequences identified for the particular clone in Table 7, as well as the corresponding plasmid vector sequences designated above.

The deposited material in the sample assigned the ATCC Deposit Number cited by reference to Tables 1, 2, 6 and 7 for any given cDNA clone also may contain one or more additional plasmids, each comprising a cDNA clone different from that given clone. Thus, deposits sharing the same ATCC Deposit Number contain at least a plasmid for each ATCC Deposit No:Z. TABLE 7 ATCC Libraries owned by Catalog Catalog Description Vector Deposit HUKA HUKB HUKC HUKD Human Uterine Cancer Lambda ZAP II LP01 HUKE HUKF HUKG HCNA HCNB Human Colon Lambda Zap II LP01 HFFA Human Fetal Brain, random Lambda Zap II LP01 primed HTWA Resting T-Cell Lambda ZAP II LP01 HBQA Early Stage Human Brain, Lambda ZAP II LP01 random primed HLMB HLMF HLMG HLMH breast lymph node CDNA Lambda ZAP II LP01 HLMI HLMJ HLMM HLMN library HCQA HCQB human colon cancer Lamda ZAP II LP01 HMEA HMEC HMED Human Microvascular Lambda ZAP II LP01 HMEE HMEF HMEG HMEI Endothelial Cells, fract. A HMEJ HMEK HMEL HUSA HUSC Human Umbilical Vein Lambda ZAP II LP01 Endothelial Cells, fract. A HLQA HLQB Hepatocellular Tumor Lambda ZAP II LP01 HHGA HHGB HHGC HHGD Hemangiopericytoma Lambda ZAP II LP01 HSDM Human Striatum Depression, Lambda ZAP II LP01 re-rescue HUSH H Umbilical Vein Endothelial Lambda ZAP II LP01 Cells, frac A, re-excision HSGS Salivary gland, subtracted Lambda ZAP II LP01 HFXA HFXB HFXC HFXD Brain frontal cortex Lambda ZAP II LP01 HFXE HFXF HFXG HFXH HPQA HPQB HPQC PERM TF274 Lambda ZAP II LP01 HFXJ HFXK Brain Frontal Cortex, re- Lambda ZAP II LP01 excision HCWA HCWB HCWC CD34 positive cells (Cord ZAP Express LP02 HCWD HCWE HCWF Blood) HCWG HCWH HCWI HCWJ HCWK HCUA HCUB HCUC CD34 depleted Buffy Coat ZAP Express LP02 (Cord Blood) HRSM A-14 cell line ZAP Express LP02 HRSA A1-CELL LINE ZAP Express LP02 HCUD HCUE HCUF HCUG CD34 depleted Buffy Coat ZAP Express LP02 HCUH HCUI (Cord Blood), re-excision HBXE HBXF HBXG H. Whole Brain #2, re- ZAP Express LP02 excision HRLM L8 cell line ZAP Express LP02 HBXA HBXB HBXC HBXD Human Whole Brain #2 - ZAP Express LP02 Oligo dT >1.5 Kb HUDA HUDB HUDC Testes ZAP Express LP02 HHTM HHTN HHTO H. hypothalamus, frac A; re- ZAP Express LP02 excision HHTL H. hypothalamus, frac A ZAP Express LP02 HASA HASD Human Adult Spleen Uni-ZAP XR LP03 HFKC HFKD HFKE HFKF Human Fetal Kidney Uni-ZAP XR LP03 HFKG HE8A HE8B HE8C HE8D Human 8 Week Whole Uni-ZAP XR LP03 HE8E HE8F HE8M HE8N Embryo HGBA HGBD HGBE HGBF Human Gall Bladder Uni-ZAP XR LP03 HGBG HGBH HGBI HLHA HLHB HLHC HLHD Human Fetal Lung III Uni-ZAP XR LP03 HLHE HLHF HLHG HLHH HLHQ HPMA HPMB HPMC HPMD Human Placenta Uni-ZAP XR LP03 HPME HPMF HPMG HPMH HPRA HPRB HPRC HPRD Human Prostate Uni-ZAP XR LP03 HSIA HSIC HSID HSIE Human Adult Small Intestine Uni-ZAP XR LP03 HTEA HTEB HTEC HTED Human Testes Uni-ZAP XR LP03 HTEE HTEF HTEG HTEH HTEI HTEJ HTEK HTPA HTPB HTPC HTPD Human Pancreas Tumor Uni-ZAP XR LP03 HTPE HTTA HTTB HTTC HTTD Human Testes Tumor Uni-ZAP XR LP03 HTTE HTTF HAPA HAPB HAPC HAPM Human Adult Pulmonary Uni-ZAP XR LP03 HETA HETB HETC HETD Human Endometrial Tumor Uni-ZAP XR LP03 HETE HETF HETG HETH HETI HHFB HHFC HHFD HHFE Human Fetal Heart Uni-ZAP XR LP03 HHFF HHFG HHFH HHFI HHPB HHPC HHPD HHPE Human Hippocampus Uni-ZAP XR LP03 HHPF HHPG HHPH HCE1 HCE2 HCE3 HCE4 Human Cerebellum Uni-ZAP XR LP03 HCE5 HCEB HCEC HCED HCEE HCEF HCEG HUVB HUVC HUVD HUVE Human Umbilical Vein, Uni-ZAP XR LP03 Endo. remake HSTA HSTB HSTC HSTD Human Skin Tumor Uni-ZAP XR LP03 HTAA HTAB HTAC HTAD Human Activated T-Cells Uni-ZAP XR LP03 HTAE HFEA HFEB HFEC Human Fetal Epithelium Uni-ZAP XR LP03 (Skin) HJPA HJPB HJPC HJPD HUMAN JURKAT Uni-ZAP XR LP03 MEMBRANE BOUND POLYSOMES HESA Human epithelioid sarcoma Uni-Zap XR LP03 HLTA HLTB HLTC HLTD Human T-Cell Lymphoma Uni-ZAP XR LP03 HLTE HLTF HFTA HFTB HFTC HFTD Human Fetal Dura Mater Uni-ZAP XR LP03 HRDA HRDB HRDC HRDD Human Rhabdomyosarcoma Uni-ZAP XR LP03 HRDE HRDF HCAA HCAB HCAC Cem cells cyclohexamide Uni-ZAP XR LP03 treated HRGA HRGB HRGC HRGD Raji Cells, cyclohexamide Uni-ZAP XR LP03 treated HSUA HSUB HSUC HSUM Supt Cells, cyclohexamide Uni-ZAP XR LP03 treated HT4A HT4C HT4D Activated T-Cells, 12 hrs. Uni-ZAP XR LP03 HE9A HE9B HE9C HE9D Nine Week Old Early Stage Uni-ZAP XR LP03 HE9E HE9F HE9G HE9H Human HE9M HE9N HATA HATB HATC HATD Human Adrenal Gland Tumor Uni-ZAP XR LP03 HATE HT5A Activated T-Cells, 24 hrs. Uni-ZAP XR LP03 HFGA HFGM Human Fetal Brain Uni-ZAP XR LP03 HNEA HNEB HNEC HNED Human Neutrophil Uni-ZAP XR LP03 HNEE HBGB HBGD Human Primary Breast Uni-ZAP XR LP03 Cancer HBNA HBNB Human Normal Breast Uni-ZAP XR LP03 HCAS Cem Cells, cyclohexamide Uni-ZAP XR LP03 treated, subtra HHPS Human Hippocampus, pBS LP03 subtracted HKCS HKCU Human Colon Cancer, pBS LP03 subtracted HRGS Raji cells, cyclohexamide pBS LP03 treated, subtracted HSUT Supt cells, cyclohexamide pBS LP03 treated, differentially expressed HT4S Activated T-Cells, 12 hrs, Uni-ZAP XR LP03 subtracted HCDA HCDB HCDC HCDD Human Chondrosarcoma Uni-ZAP XR LP03 HCDE HOAA HOAB HOAC Human Osteosarcoma Uni-ZAP XR LP03 HTLA HTLB HTLC HTLD Human adult testis, large Uni-ZAP XR LP03 HTLE HTLF inserts HLMA HLMC HLMD Breast Lymph node cDNA Uni-ZAP XR LP03 library H6EA H6EB H6EC HL-60, PMA 4 H Uni-ZAP XR LP03 HTXA HTXB HTXC HTXD Activated T-Cell Uni-ZAP XR LP03 HTXE HTXF HTXG HTXH (12 hs)/Thiouridine labelledEco HNFA HNFB HNFC HNFD Human Neutrophil, Activated Uni-ZAP XR LP03 HNFE HNFF HNFG HNFH HNFJ HTOB HTOC HUMAN TONSILS, Uni-ZAP XR LP03 FRACTION 2 HMGB Human OB MG63 control Uni-ZAP XR LP03 fraction I HOPB Human OB HOS control Uni-ZAP XR LP03 fraction I HORB Human OB HOS treated (10 nM Uni-ZAP XR LP03 E2) fraction I HSVA HSVB HSVC Human Chronic Synovitis Uni-ZAP XR LP03 HROA HUMAN STOMACH Uni-ZAP XR LP03 HBJA HBJB HBJC HBJD HUMAN B CELL Uni-ZAP XR LP03 HBJE HBJF HBJG HBJH LYMPHOMA HBJI HBJJ HBJK HCRA HCRB HCRC human corpus colosum Uni-ZAP XR LP03 HODA HODB HODC HODD human ovarian cancer Uni-ZAP XR LP03 HDSA Dermatofibrosarcoma Uni-ZAP XR LP03 Protuberance HMWA HMWB HMWC Bone Marrow Cell Line Uni-ZAP XR LP03 HMWD HMWE HMWF (RS4; 11) HMWG HMWH HMWI HMWJ HSOA stomach cancer (human) Uni-ZAP XR LP03 HERA SKIN Uni-ZAP XR LP03 HMDA Brain-medulloblastoma Uni-ZAP XR LP03 HGLA HGLB HGLD Glioblastoma Uni-ZAP XR LP03 HEAA H. Atrophic Endometrium Uni-ZAP XR LP03 HBCA HBCB H. Lymph node breast Cancer Uni-ZAP XR LP03 HPWT Human Prostate BPH, re- Uni-ZAP XR LP03 excision HFVG HFVH HFVI Fetal Liver, subtraction II pBS LP03 HNFI Human Neutrophils, pBS LP03 Activated, re-excision HBMB HBMC HBMD Human Bone Marrow, re- pBS LP03 excision HKML HKMM HKMN H. Kidney Medulla, re- pBS LP03 excision HKIX HKIY H. Kidney Cortex, subtracted pBS LP03 HADT H. Amygdala Depression, pBS LP03 subtracted H6AS Hl-60, untreated, subtracted Uni-ZAP XR LP03 H6ES HL-60, PMA 4 H, subtracted Uni-ZAP XR LP03 H6BS HL-60, RA 4 h, Subtracted Uni-ZAP XR LP03 H6CS HL-60, PMA 1 d, subtracted Uni-ZAP XR LP03 HTXJ HTXK Activated T- Uni-ZAP XR LP03 cell(12 h)/Thiouridine-re- excision HMSA HMSB HMSC HMSD Monocyte activated Uni-ZAP XR LP03 HMSE HMSF HMSG HMSH HMSI HMSJ HMSK HAGA HAGB HAGC HAGD Human Amygdala Uni-ZAP XR LP03 HAGE HAGF HSRA HSRB HSRE STROMAL - Uni-ZAP XR LP03 OSTEOCLASTOMA HSRD HSRF HSRG HSRH Human Osteoclastoma Uni-ZAP XR LP03 Stromal Cells - unamplified HSQA HSQB HSQC HSQD Stromal cell TF274 Uni-ZAP XR LP03 HSQE HSQF HSQG HSKA HSKB HSKC HSKD Smooth muscle, serum treated Uni-ZAP XR LP03 HSKE HSKF HSKZ HSLA HSLB HSLC HSLD Smooth muscle, control Uni-ZAP XR LP03 HSLE HSLF HSLG HSDA HSDD HSDE HSDF Spinal cord Uni-ZAP XR LP03 HSDG HSDH HPWS Prostate-BPH subtracted II pBS LP03 HSKW HSKX HSKY Smooth Muscle-HASTE pBS LP03 normalized HFPB HFPC HFPD H. Frontal cortex, epileptic; re- Uni-ZAP XR LP03 excision HSDI HSDJ HSDK Spinal Cord, re-excision Uni-ZAP XR LP03 HSKN HSKO Smooth Muscle Serum pBS LP03 Treated, Norm HSKG HSKH HSKI Smooth muscle, serum pBS LP03 induced, re-exc HFCA HFCB HFCC HFCD Human Fetal Brain Uni-ZAP XR LP04 HFCE HFCF HPTA HPTB HPTD Human Pituitary Uni-ZAP XR LP04 HTHB HTHC HTHD Human Thymus Uni-ZAP XR LP04 HE6B HE6C HE6D HE6E Human Whole Six Week Old Uni-ZAP XR LP04 HE6F HE6G HE6S Embryo HSSA HSSB HSSC HSSD Human Synovial Sarcoma Uni-ZAP XR LP04 HSSE HSSF HSSG HSSH HSSI HSSJ HSSK HE7T 7 Week Old Early Stage Uni-ZAP XR LP04 Human, subtracted HEPA HEPB HEPC Human Epididymus Uni-ZAP XR LP04 HSNA HSNB HSNC HSNM Human Synovium Uni-ZAP XR LP04 HSNN HPFB HPFC HPFD HPFE Human Prostate Cancer, Uni-ZAP XR LP04 Stage C fraction HE2A HE2D HE2E HE2H 12 Week Old Early Stage Uni-ZAP XR LP04 HE2I HE2M HE2N HE2O Human HE2B HE2C HE2F HE2G 12 Week Old Early Stage Uni-ZAP XR LP04 HE2P HE2Q Human, II HPTS HPTT HPTU Human Pituitary, subtracted Uni-ZAP XR LP04 HAUA HAUB HAUC Amniotic Cells - TNF Uni-ZAP XR LP04 induced HAQA HAQB HAQC HAQD Amniotic Cells - Primary Uni-ZAP XR LP04 Culture HWTA HWTB HWTC wilm's tumor Uni-ZAP XR LP04 HBSD Bone Cancer, re-excision Uni-ZAP XR LP04 HSGB Salivary gland, re-excision Uni-ZAP XR LP04 HSJA HSJB HSJC Smooth muscle-ILb induced Uni-ZAP XR LP04 HSXA HSXB HSXC HSXD Human Substantia Nigra Uni-ZAP XR LP04 HSHA HSHB HSHC Smooth muscle, IL1b induced Uni-ZAP XR LP04 HOUA HOUB HOUC HOUD Adipocytes Uni-ZAP XR LP04 HOUE HPWA HPWB HPWC Prostate BPH Uni-ZAP XR LP04 HPWD HPWE HELA HELB HELC HELD Endothelial cells-control Uni-ZAP XR LP04 HELE HELF HELG HELH HEMA HEMB HEMC Endothelial-induced Uni-ZAP XR LP04 HEMD HEME HEMF HEMG HEMH HBIA HBIB HBIC Human Brain, Striatum Uni-ZAP XR LP04 HHSA HHSB HHSC HHSD Human Uni-ZAP XR LP04 HHSE Hypothalmus, Schizophrenia HNGA HNGB HNGC HNGD neutrophils control Uni-ZAP XR LP04 HNGE HNGF HNGG HNGH HNGI HNGJ HNHA HNHB HNHC HNHD Neutrophils IL-1 and LPS Uni-ZAP XR LP04 HNHE HNHF HNHG HNHH induced HNHI HNHJ HSDB HSDC STRIATUM DEPRESSION Uni-ZAP XR LP04 HHPT Hypothalamus Uni-ZAP XR LP04 HSAT HSAU HSAV HSAW Anergic T-cell Uni-ZAP XR LP04 HSAX HSAY HSAZ HBMS HBMT HBMU Bone marrow Uni-ZAP XR LP04 HBMV HBMW HBMX HOEA HOEB HOEC HOED Osteoblasts Uni-ZAP XR LP04 HOEE HOEF HOEJ HAIA HAIB HAIC HAID Epithelial-TNFa and INF Uni-ZAP XR LP04 HAIE HAIF induced HTGA HTGB HTGC HTGD Apoptotic T-cell Uni-ZAP XR LP04 HMCA HMCB HMCC Macrophage-oxLDL Uni-ZAP XR LP04 HMCD HMCE HMAA HMAB HMAC Macrophage (GM-CSF Uni-ZAP XR LP04 HMAD HMAE HMAF treated) HMAG HPHA Normal Prostate Uni-ZAP XR LP04 HPIA HPIB HPIC LNCAP prostate cell line Uni-ZAP XR LP04 HPJA HPJB HPJC PC3 Prostate cell line Uni-ZAP XR LP04 HOSE HOSF HOSG Human Osteoclastoma, re- Uni-ZAP XR LP04 excision HTGE HTGF Apoptotic T-cell, re-excision Uni-ZAP XR LP04 HMAJ HMAK H Macrophage (GM-CSF Uni-ZAP XR LP04 treated), re-excision HACB HACC HACD Human Adipose Tissue, re- Uni-ZAP XR LP04 excision HFPA H. Frontal Cortex, Epileptic Uni-ZAP XR LP04 HFAA HFAB HFAC HFAD Alzheimer's, spongy change Uni-ZAP XR LP04 HFAE HFAM Frontal Lobe, Dementia Uni-ZAP XR LP04 HMIA HMIB HMIC Human Manic Depression Uni-ZAP XR LP04 Tissue HTSA HTSE HTSF HTSG Human Thymus pBS LP05 HTSH HPBA HPBB HPBC HPBD Human Pineal Gland pBS LP05 HPBE HSAA HSAB HSAC HSA 172 Cells pBS LP05 HSBA HSBB HSBC HSBM HSC172 cells HJAA HJAB HJAC HJAD Jurkat T-cell G1 phase pBS LP05 HJBA HJBB HJBC HJBD Jurkat T-Cell, S phase pBS LP05 HAFA HAFB Aorta endothelial cells + TNF-a pBS LP05 HAWA HAWB HAWC Human White Adipose pBS LP05 HTNA HTNB Human Thyroid pBS LP05 HONA Normal Ovary, pBS LP05 Premenopausal HARA HARB Human Adult Retina pBS LP05 HLJA HLJB Human Lung pCMVSport 1 LP06 HOFM HOFN HOFO H. Ovarian Tumor, II, pCMVSport 2.0 LP07 OV5232 HOGA HOGB HOGC OV 10-3-95 pCMVSport 2.0 LP07 HCGL CD34+cells, II pCMVSport 2.0 LP07 HDLA Hodgkin's Lymphoma I pCMVSport 2.0 LP07 HDTA HDTB HDTC HDTD Hodgkin's Lymphoma II pCMVSport 2.0 LP07 HDTE HKAA HKAB HKAC HKAD Keratinocyte pCMVSport 2.0 LP07 HKAE HKAF HKAG HKAH HCIM CAPFINDER, Crohn's pCMVSport 2.0 LP07 Disease, lib 2 HKAL Keratinocyte, lib 2 pCMVSport 2.0 LP07 HKAT Keratinocyte, lib 3 pCMVSport 2.0 LP07 HNDA Nasal polyps pCMVSport 2.0 LP07 HDRA H. Primary Dendritic Cells, lib 3 pCMVSport 2.0 LP07 HOHA HOHB HOHC Human Osteoblasts II pCMVSport 2.0 LP07 HLDA HLDB HLDC Liver, Hepatoma pCMVSport 3.0 LP08 HLDN HLDO HLDP Human Liver, normal pCMVSport 3.0 LP08 HMTA pBMC stimulated w/poly I/C pCMVSport 3.0 LP08 HNTA NTERA2, control pCMVSport 3.0 LP08 HDPA HDPB HDPC HDPD Primary Dendritic Cells, lib 1 pCMVSport 3.0 LP08 HDPF HDPG HDPH HDPI HDPJ HDPK HDPM HDPN HDPO HDPP Primary Dendritic cells, frac 2 pCMVSport 3.0 LP08 HMUA HMUB HMUC Myoloid Progenitor Cell Line pCMVSport 3.0 LP08 HHEA HHEB HHEC HHED T Cell helper I pCMVSport 3.0 LP08 HHEM HHEN HHEO HHEP T cell helper II pCMVSport 3.0 LP08 HEQA HEQB HEQC Human endometrial stromal pCMVSport 3.0 LP08 cells HJMA HJMB Human endometrial stromal pCMVSport 3.0 LP08 cells-treated with progesterone HSWA HSWB HSWC Human endometrial stromal pCMVSport 3.0 LP08 cells-treated with estradiol HSYA HSYB HSYC Human Thymus Stromal Cells pCMVSport 3.0 LP08 HLWA HLWB HLWC Human Placenta pCMVSport 3.0 LP08 HRAA HRAB HRAC Rejected Kidney, lib 4 pCMVSport 3.0 LP08 HMTM PCR, pBMC I/C treated PCRII LP09 HMJA H. Meniingima, M6 pSport 1 LP10 HMKA HMKB HMKC H. Meningima, M1 pSport 1 LP10 HMKD HMKE HUSG HUSI Human umbilical vein pSport 1 LP10 endothelial cells, IL-4 induced HUSX HUSY Human Umbilical Vein pSport 1 LP10 Endothelial Cells, uninduced HOFA Ovarian Tumor I, OV5232 pSport 1 LP10 HCFA HCFB HCFC HCFD T-Cell PHA 16 hrs pSport 1 LP10 HCFL HCFM HCFN HCFO T-Cell PHA 24 hrs pSport 1 LP10 HADA HADC HADD HADE Human Adipose pSport 1 LP10 HADF HADG HOVA HOVB HOVC Human Ovary pSport 1 LP10 HTWB HTWC HTWD Resting T-Cell Library, II pSport 1 LP10 HTWE HTWF HMMA Spleen metastic melanoma pSport 1 LP10 HLYA HLYB HLYC HLYD Spleen, Chronic lymphocytic pSport 1 LP10 HLYE leukemia HCGA CD34+ cell, I pSport 1 LP10 HEOM HEON Human Eosinophils pSport 1 LP10 HTDA Human Tonsil, Lib 3 pSport 1 LP10 HSPA Salivary Gland, Lib 2 pSport 1 LP10 HCHA HCHB HCHC Breast Cancer cell line, MDA pSport 1 LP10 36 HCHM HCHN Breast Cancer Cell line, pSport 1 LP10 angiogenic HCIA Crohn's Disease pSport 1 LP10 HDAA HDAB HDAC HEL cell line pSport 1 LP10 HABA Human Astrocyte pSport 1 LP10 HUFA HUFB HUFC Ulcerative Colitis pSport 1 LP10 HNTM NTERA2 + retinoic acid, 14 pSport 1 LP10 days HDQA Primary Dendritic pSport 1 LP10 cells, CapFinder2, frac 1 HDQM Primary Dendritic Cells, pSport 1 LP10 CapFinder, frac 2 HLDX Human Liver, pSport 1 LP10 normal, CapFinder HULA HULB HULC Human Dermal Endothelial pSport1 LP10 Cells, untreated HUMA Human Dermal Endothelial pSport1 LP10 cells, treated HCJA Human Stromal Endometrial pSport1 LP10 fibroblasts, untreated HCJM Human Stromal endometrial pSport1 LP10 fibroblasts, treated w/ estradiol HEDA Human Stromal endometrial pSport1 LP10 fibroblasts, treated with progesterone HFNA Human ovary tumor cell pSport1 LP10 OV350721 HKGA HKGB HKGC HKGD Merkel Cells pSport1 LP10 HISA HISB HISC Pancreas Islet Cell Tumor pSport1 LP10 HLSA Skin, burned pSport1 LP10 HBZA Prostate, BPH, Lib 2 pSport 1 LP10 HBZS Prostate BPH, Lib 2, pSport 1 LP10 subtracted HFIA HFIB HFIC Synovial Fibroblasts (control) pSport 1 LP10 HFIH HFII HFIJ Synovial hypoxia pSport 1 LP10 HFIT HFIU HFIV Synovial IL-1/TNF stimulated pSport 1 LP10 HGCA Messangial cell, frac 1 pSport1 LP10 HMVA HMVB HMVC Bone Marrow Stromal Cell, pSport1 LP10 untreated HFIX HFIY HFIZ Synovial Fibroblasts pSport1 LP10 (Il1/TNF), subt HFOX HFOY HFOZ Synovial hypoxia-RSF pSport1 LP10 subtracted HMQA HMQB HMQC Human Activated Monocytes Uni-ZAP XR LP11 HMQD HLIA HLIB HLIC Human Liver pCMVSport 1 LP012 HHBA HHBB HHBC HHBD Human Heart pCMVSport 1 LP012 HHBE HBBA HBBB Human Brain pCMVSport 1 LP012 HLJA HLJB HLJC HLJD Human Lung pCMVSport 1 LP012 HLJE HOGA HOGB HOGC Ovarian Tumor pCMVSport 2.0 LP012 HTJM Human Tonsils, Lib 2 pCMVSport 2.0 LP012 HAMF HAMG KMH2 pCMVSport 3.0 LP012 HAJA HAJB HAJC L428 pCMVSport 3.0 LP012 HWBA HWBB HWBC Dendritic cells, pooled pCMVSport 3.0 LP012 HWBD HWBE HWAA HWAB HWAC Human Bone Marrow, pCMVSport 3.0 LP012 HWAD HWAE treated HYAA HYAB HYAC B Cell lymphoma pCMVSport 3.0 LP012 HWHG HWHH HWHI Healing groin wound, 6.5 pCMVSport 3.0 LP012 hours post incision HWHP HWHQ HWHR Healing groin wound; 7.5 pCMVSport 3.0 LP012 hours post incision HARM Healing groin wound - zero hr pCMVSport 3.0 LP012 post-incision (control) HBIM Olfactory epithelium; pCMVSport 3.0 LP012 nasalcavity HWDA Healing Abdomen wound; pCMVSport 3.0 LP012 70&90 min post incision HWEA Healing Abdomen Wound; 15 pCMVSport 3.0 LP012 days post incision HWJA Healing Abdomen pCMVSport 3.0 LP012 Wound; 21&29 days HNAL Human Tongue, frac 2 pSport1 LP012 HMJA H. Meniingima, M6 pSport1 LP012 HMKA HMKB HMKC H. Meningima, M1 pSport1 LP012 HMKD HMKE HOFA Ovarian Tumor I, OV5232 pSport1 LP012 HCFA HCFB HCFC HCFD T-Cell PHA 16 hrs pSport1 LP012 HCFL HCFM HCFN HCFO T-Cell PHA 24 hrs pSport1 LP012 HMMA HMMB HMMC Spleen metastic melanoma pSport1 LP012 HTDA Human Tonsil, Lib 3 pSport1 LP012 HDBA Human Fetal Thymus pSport1 LP012 HDUA Pericardium pSport1 LP012 HBZA Prostate, BPH, Lib 2 pSport1 LP012 HWCA Larynx tumor pSport1 LP012 HWKA Normal lung pSport1 LP012 HSMB Bone marrow stroma, treated pSport1 LP012 HBHM Normal trachea pSport1 LP012 HLFC Human Larynx pSport1 LP012 HLRB Siebben Polyposis pSport1 LP012 HNIA Mammary Gland pSport1 LP012 HNJB Palate carcinoma pSport1 LP012 HNKA Palate normal pSport1 LP012 HMZA Pharynx carcinoma pSport1 LP012 HABG Cheek Carcinoma pSport1 LP012 HMZM Pharynx Carcinoma pSport1 LP012 HDRM Larynx Carcinoma pSport1 LP012 HVAA Pancreas normal PCA4 No pSport1 LP012 HICA Tongue carcinoma pSport1 LP012 HUKA HUKB HUKC HUKD Human Uterine Cancer Lambda ZAP II LP013 HUKE HFFA Human Fetal Brain, random Lambda ZAP II LP013 primed HTUA Activated T-cell labeled with Lambda ZAP II LP013 4-thioluri HBQA Early Stage Human Brain, Lambda ZAP II LP013 random primed HMEB Human microvascular Lambda ZAP II LP013 Endothelial cells, fract. B HUSH Human Umbilical Vein Lambda ZAP II LP013 Endothelial cells, fract. A, re- excision HLQC HLQD Hepatocellular tumor, re- Lambda ZAP II LP013 excision HTWJ HTWK HTWL Resting T-cell, re-excision Lambda ZAP II LP013 HF6S Human Whole 6 week Old pBluescript LP013 Embryo (II), subt HHPS Human Hippocampus, pBluescript LP013 subtracted HL1S LNCAP, differential pBluescript LP013 expression HLHS HLHT Early Stage Human Lung, pBluescript LP013 Subtracted HSUS Supt cells, cyclohexamide pBluescript LP013 treated, subtracted HSUT Supt cells, cyclohexamide pBluescript LP013 treated, differentially expressed HSDS H. Striatum Depression, pBluescript LP013 subtracted HPTZ Human Pituitary, Subtracted pBluescript LP013 VII HSDX H. Striatum Depression, subt pBluescript LP013 II HSDZ H. Striatum Depression, subt pBluescript LP013 HPBA HPBB HPBC HPBD Human Pineal Gland pBluescript SK− LP013 HPBE HRTA Colorectal Tumor pBluescript SK− LP013 HSBA HSBB HSBC HSBM HSC172 cells pBluescript SK− LP013 HJAA HJAB HJAC HJAD Jurkat T-cell G1 phase pBluescript SK− LP013 HJBA HJBB HJBC HJBD Jurkat T-cell, S1 phase pBluescript SK− LP013 HTNA HTNB Human Thyroid pBluescript SK− LP013 HAHA HAHB Human Adult Heart Uni-ZAP XR LP013 HE6A Whole 6 week Old Embryo Uni-ZAP XR LP013 HFCA HFCB HFCC HFCD Human Fetal Brain Uni-ZAP XR LP013 HFCE HFKC HFKD HFKE HFKF Human Fetal Kidney Uni-ZAP XR LP013 HFKG HGBA HGBD HGBE HGBF Human Gall Bladder Uni-ZAP XR LP013 HGBG HPRA HPRB HPRC HPRD Human Prostate Uni-ZAP XR LP013 HTEA HTEB HTEC HTED Human Testes Uni-ZAP XR LP013 HTEE HTTA HTTB HTTC HTTD Human Testes Tumor Uni-ZAP XR LP013 HTTE HYBA HYBB Human Fetal Bone Uni-ZAP XR LP013 HFLA Human Fetal Liver Uni-ZAP XR LP013 HHFB HHFC HHFD HHFE Human Fetal Heart Uni-ZAP XR LP013 HHFF HUVB HUVC HUVD HUVE Human Umbilical Vein, End. Uni-ZAP XR LP013 remake HTHB HTHC HTHD Human Thymus Uni-ZAP XR LP013 HSTA HSTB HSTC HSTD Human Skin Tumor Uni-ZAP XR LP013 HTAA HTAB HTAC HTAD Human Activated T-cells Uni-ZAP XR LP013 HTAE HFEA HFEB HFEC Human Fetal Epithelium Uni-ZAP XR LP013 (skin) HJPA HJPB HJPC HJPD Human Jurkat Membrane Uni-ZAP XR LP013 Bound Polysomes HESA Human Epithelioid Sarcoma Uni-ZAP XR LP013 HALS Human Adult Liver, Uni-ZAP XR LP013 Subtracted HFTA HFTB HFTC HFTD Human Fetal Dura Mater Uni-ZAP XR LP013 HCAA HCAB HCAC Cem cells, cyclohexamide Uni-ZAP XR LP013 treated HRGA HRGB HRGC HRGD Raji Cells, cyclohexamide Uni-ZAP XR LP013 treated HE9A HE9B HE9C HE9D Nine Week Old Early Stage Uni-ZAP XR LP013 HE9E Human HSFA Human Fibrosarcoma Uni-ZAP XR LP013 HATA HATB HATC HATD Human Adrenal Gland Tumor Uni-ZAP XR LP013 HATE HTRA Human Trachea Tumor Uni-ZAP XR LP013 HE2A HE2D HE2E HE2H 12 Week Old Early Stage Uni-ZAP XR LP013 HE2I Human HE2B HE2C HE2F HE2G 12 Week Old Early Stage Uni-ZAP XR LP013 HE2P Human, II HNEA HNEB HNEC HNED Human Neutrophil Uni-ZAP XR LP013 HNEE HBGA Human Primary Breast Uni-ZAP XR LP013 Cancer HPTS HPTT HPTU Human Pituitary, subtracted Uni-ZAP XR LP013 HMQA HMQB HMQC Human Activated Monocytes Uni-ZAP XR LP013 HMQD HOAA HOAB HOAC Human Osteosarcoma Uni-ZAP XR LP013 HTOA HTOD HTOE HTOF human tonsils Uni-ZAP XR LP013 HTOG HMGB Human OB MG63 control Uni-ZAP XR LP013 fraction I HOPB Human OB HOS control Uni-ZAP XR LP013 fraction I HOQB Human OB HOS treated (1 nM Uni-ZAP XR LP013 E2) fraction I HAUA HAUB HAUC Amniotic Cells - TNF Uni-ZAP XR LP013 induced HAQA HAQB HAQC HAQD Amniotic Cells - Primary Uni-ZAP XR LP013 Culture HROA HROC HUMAN STOMACH Uni-ZAP XR LP013 HBJA HBJB HBJC HBJD HUMAN B CELL Uni-ZAP XR LP013 HBJE LYMPHOMA HODA HODB HODC HODD human ovarian cancer Uni-ZAP XR LP013 HCPA Corpus Callosum Uni-ZAP XR LP013 HSOA stomach cancer (human) Uni-ZAP XR LP013 HERA SKIN Uni-ZAP XR LP013 HMDA Brain-medulloblastoma Uni-ZAP XR LP013 HGLA HGLB HGLD Glioblastoma Uni-ZAP XR LP013 HWTA HWTB HWTC wilm's tumor Uni-ZAP XR LP013 HEAA H. Atrophic Endometrium Uni-ZAP XR LP013 HAPN HAPO HAPP HAPQ Human Adult Pulmonary; re- Uni-ZAP XR LP013 HAPR excision HLTG HLTH Human T-cell lymphoma; re- Uni-ZAP XR LP013 excision HAHC HAHD HAHE Human Adult Heart; re- Uni-ZAP XR LP013 excision HAGA HAGB HAGC HAGD Human Amygdala Uni-ZAP XR LP013 HAGE HSJA HSJB HSJC Smooth muscle-ILb induced Uni-ZAP XR LP013 HSHA HSHB HSHC Smooth muscle, IL1b induced Uni-ZAP XR LP013 HPWA HPWB HPWC Prostate BPH Uni-ZAP XR LP013 HPWD HPWE HPIA HPIB HPIC LNCAP prostate cell line Uni-ZAP XR LP013 HPJA HPJB HPJC PC3 Prostate cell line Uni-ZAP XR LP013 HBTA Bone Marrow Stroma, Uni-ZAP XR LP013 TNF&LPS ind HMCF HMCG HMCH HMCI Macrophage-oxLDL; re- Uni-ZAP XR LP013 HMCJ excision HAGG HAGH HAGI Human Amygdala; re-excision Uni-ZAP XR LP013 HACA H. Adipose Tissue Uni-ZAP XR LP013 HKFB K562 + PMA (36 hrs), re- ZAP Express LP013 excision HCWT HCWU HCWV CD34 positive cells (cord ZAP Express LP013 blood), re-ex HBWA Whole brain ZAP Express LP013 HBXA HBXB HBXC HBXD Human Whole Brain #2 - ZAP Express LP013 Oligo dT > 1.5 Kb HAVM Temporal cortex-Alzheizmer pT-Adv LP014 HAVT Hippocampus, Alzheimer pT-Adv LP014 Subtracted HHAS CHME Cell Line Uni-ZAP XR LP014 HAJR Larynx normal pSport 1 LP014 HWLE HWLF HWLG Colon Normal pSport 1 LP014 HWLH HCRM HCRN HCRO Colon Carcinoma pSport 1 LP014 HWLI HWLJ HWLK Colon Normal pSport 1 LP014 HWLQ HWLR HWLS Colon Tumor pSport 1 LP014 HWLT HBFM Gastrocnemius Muscle pSport 1 LP014 HBOD HBOE Quadriceps Muscle pSport 1 LP014 HBKD HBKE Soleus Muscle pSport 1 LP014 HCCM Pancreatic Langerhans pSport 1 LP014 HWGA Larynx carcinoma pSport 1 LP014 HWGM HWGN Larynx carcinoma pSport 1 LP014 HWLA HWLB HWLC Normal colon pSport 1 LP014 HWLM HWLN Colon Tumor pSport 1 LP014 HVAM HVAN HVAO Pancreas Tumor pSport 1 LP014 HWGQ Larynx carcinoma pSport 1 LP014 HAQM HAQN Salivary Gland pSport 1 LP014 HASM Stomach; normal pSport 1 LP014 HBCM Uterus; normal pSport 1 LP014 HCDM Testis; normal pSport 1 LP014 HDJM Brain; normal pSport 1 LP014 HEFM Adrenal Gland, normal pSport 1 LP014 HBAA Rectum normal pSport 1 LP014 HFDM Rectum tumour pSport 1 LP014 HGAM Colon, normal pSport 1 LP014 HHMM Colon, tumour pSport 1 LP014 HCLB HCLC Human Lung Cancer Lambda Zap II LP015 HRLA L1 Cell line ZAP Express LP015 HHAM Hypothalamus, Alzheimer's pCMVSport 3.0 LP015 HKBA Ku 812F Basophils Line pSport 1 LP015 HS2S Saos2, Dexamethosome pSport 1 LP016 Treated HA5A Lung Carcinoma A549 pSport 1 LP016 TNFalpha activated HTFM TF-1 Cell Line GM-CSF pSport 1 LP016 Treated HYAS Thyroid Tumour pSport 1 LP016 HUTS Larynx Normal pSport 1 LP016 HXOA Larynx Tumor pSport 1 LP016 HEAH Ea.hy.926 cell line pSport 1 LP016 HINA Adenocarcinoma Human pSport 1 LP016 HRMA Lung Mesothelium pSport 1 LP016 HLCL Human Pre-Differentiated Uni-Zap XR LP017 Adipocytes HS2A Saos2 Cells pSport 1 LP020 HS2I Saos2 Cells; Vitamin D3 pSport 1 LP020 Treated HUCM CHME Cell Line, untreated pSport 1 LP020 HEPN Aryepiglottis Normal pSport 1 LP020 HPSN Sinus Piniformis Tumour pSport 1 LP020 HNSA Stomach Normal pSport 1 LP020 HNSM Stomach Tumour pSport 1 LP020 HNLA Liver Normal Met5No pSport 1 LP020 HUTA Liver Tumour Met 5 Tu pSport 1 LP020 HOCN Colon Normal pSport 1 LP020 HOCT Colon Tumor pSport 1 LP020 HTNT Tongue Tumour pSport 1 LP020 HLXN Larynx Normal pSport 1 LP020 HLXT Larynx Tumour pSport 1 LP020 HTYN Thymus pSport 1 LP020 HPLN Placenta pSport 1 LP020 HTNG Tongue Normal pSport 1 LP020 HZAA Thyroid Normal (SDCA2 No) pSport 1 LP020 HWES Thyroid Thyroiditis pSport 1 LP020 HFHD Ficolled Human Stromal pTrip1Ex2 LP021 Cells, 5Fu treated HFHM, HFHN Ficolled Human Stromal pTrip1Ex2 LP021 Cells, Untreated HPCI Hep G2 Cells, lambda library lambda Zap-CMV LP021 XR HBCA, HBCB, HBCC H. Lymph node breast Cancer Uni-ZAP XR LP021 HCOK Chondrocytes pSPORT1 LP022 HDCA, HDCB, HDCC Dendritic Cells From CD34 pSPORT1 LP022 Cells HDMA, HDMB CD40 activated monocyte pSPORT1 LP022 dendritic cells HDDM, HDDN, HDDO LPS activated derived pSPORT1 LP022 dendritic cells HPCR Hep G2 Cells, PCR library lambda Zap-CMV LP022 XR HAAA, HAAB, HAAC Lung, Cancer (4005313A3): pSPORT1 LP022 Invasive Poorly Differentiated Lung Adenocarcinoma HIPA, HIPB, HIPC Lung, Cancer (4005163 B7): pSPORT1 LP022 Invasive, Poorly Diff. Adenocarcinoma, Metastatic HOOH, HOOI Ovary, Cancer: (4004562 B6) pSPORT1 LP022 Papillary Serous Cystic Neoplasm, Low Malignant Pot HIDA Lung, Normal: (4005313 B1) pSPORT1 LP022 HUJA, HUJB, HUJC, HUJD, HUJE B-Cells pCMVSport 3.0 LP022 HNOA, HNOB, HNOC, HNOD Ovary, Normal: (9805C040R) pSPORT1 LP022 HNLM Lung, Normal: (4005313 B1) pSPORT1 LP022 HSCL Stromal Cells pSPORT1 LP022 HAAX Lung, Cancer: (4005313 A3) pSPORT1 LP022 Invasive Poorly-differentiated Metastatic lung adenocarcinoma HUUA, HUUB, HUUC, HUUD B-cells (unstimulated) pTrip1Ex2 LP022 HWWA, HWWB, HWWC, HWWD, B-cells (stimulated) pSPORT1 LP022 HWWE, HWWF, HWWG HCCC Colon, Cancer: (9808C064R) pCMVSport 3.0 LP023 HPDO HPDP HPDQ HPDR Ovary, Cancer (9809C332): pSport 1 LP023 HPD Poorly differentiated adenocarcinoma HPCO HPCP HPCQ HPCT Ovary, Cancer (15395A1F): pSport 1 LP023 Grade II Papillary Carcinoma HOCM HOCO HOCP HOCQ Ovary, Cancer: (15799A1F) pSport 1 LP023 Poorly differentiated carcinoma HCBM HCBN HCBO Breast, Cancer: (4004943 A5) pSport 1 LP023 HNBT HNBU HNBV Breast, Normal: (4005522B2) pSport 1 LP023 HBCP HBCQ Breast, Cancer: (4005522 A2) pSport 1 LP023 HBCJ Breast, Cancer: (9806C012R) pSport 1 LP023 HSAM HSAN Stromal cells 3.88 pSport 1 LP023 HVCA HVCB HVCC HVCD Ovary, Cancer: (4004332 A2) pSport 1 LP023 HSCK HSEN HSEO Stromal cells (HBM3.18) pSport 1 LP023 HSCP HSCQ stromal cell clone 2.5 pSport 1 LP023 HUXA Breast Cancer: (4005385 A2) pSport 1 LP023 HCOM HCON HCOO HCOP Ovary, Cancer (4004650 A3): pSport 1 LP023 HCOQ Well-Differentiated Micropapillary Serous Carcinoma HBNM Breast, Cancer: (9802C020E) pSport 1 LP023 HVVA HVVB HVVC HVVD Human Bone Marrow, treated pSport 1 LP023 HVVE

Two nonlimiting examples are provided below for isolating a particular clone from the deposited sample of plasmid cDNAs cited for that clone in Table 7. First, a plasmid is directly isolated by screening the clones using a polynucleotide probe corresponding to the nucleotide sequence of SEQ ID NO:X.

Particularly, a specific polynucleotide with 30-40 nucleotides is synthesized using an Applied Biosystems DNA synthesizer according to the sequence reported. The oligonucleotide is labeled, for instance, with ³²P-γ-ATP using T4 polynucleotide kinase and purified according to routine methods. (E.g., Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring, N.Y. (1982)). The plasmid mixture is transformed into a suitable host, as indicated above (such as XL-1 Blue (Stratagene)) using techniques known to those of skill in the art, such as those provided by the vector supplier or in related publications or patents cited above. The transformants are plated on 1.5% agar plates (containing the appropriate selection agent, e.g., ampicillin) to a density of about 150 transformants (colonies) per plate. These plates are screened using Nylon membranes according to routine methods for bacterial colony screening (e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edit., (1989), Cold Spring Harbor Laboratory Press, pages 1.93 to 1.104), or other techniques known to those of skill in the art.

Alternatively, two primers of 17-20 nucleotides derived from both ends of the nucleotide sequence of SEQ ID NO:X are synthesized and used to amplify the desired cDNA using the deposited cDNA plasmid as a template. The polymerase chain reaction is carried out under routine conditions, for instance, in 25 μl of reaction mixture with 0.5 ug of the above cDNA template. A convenient reaction mixture is 1.5-5 mM MgCl₂, 0.01% (w/v) gelatin, 20 μM each of dATP, dCTP, dGTP, dTTP, 25 pmol of each primer and 0.25 Unit of Taq polymerase. Thirty five cycles of PCR (denaturation at 94° C. for 1 min; annealing at 55° C. for 1 min; elongation at 72° C. for 1 min) are performed with a Perkin-Elmer Cetus automated thermal cycler. The amplified product is analyzed by agarose gel electrophoresis and the DNA band with expected molecular weight is excised and purified. The PCR product is verified to be the selected sequence by subcloning and sequencing the DNA product.

Several methods are available for the identification of the 5′ or 3′ non-coding portions of a gene which may not be present in the deposited clone. These methods include but are not limited to, filter probing, clone enrichment using specific probes, and protocols similar or identical to 5′ and 3′ “RACE” protocols which are well known in the art. For instance, a method similar to 5′ RACE is available for generating the missing 5′ end of a desired full-length transcript. (Fromont-Racine et al., Nucleic Acids Res. 21(7):1683-1684 (1993)).

Briefly, a specific RNA oligonucleotide is ligated to the 5′ ends of a population of RNA presumably containing full-length gene RNA transcripts. A primer set containing a primer specific to the ligated RNA oligonucleotide and a primer specific to a known sequence of the gene of interest is used to PCR amplify the 5′ portion of the desired full-length gene. This amplified product may then be sequenced and used to generate the full length gene.

This above method starts with total RNA isolated from the desired source, although poly-A+ RNA can be used. The RNA preparation can then be treated with phosphatase if necessary to eliminate 5′ phosphate groups on degraded or damaged RNA which may interfere with the later RNA ligase step. The phosphatase should then be inactivated and the RNA treated with tobacco acid pyrophosphatase in order to remove the cap structure present at the 5′ ends of messenger RNAs. This reaction leaves a 5′ phosphate group at the 5′ end of the cap cleaved RNA which can then be ligated to an RNA oligonucleotide using T4 RNA ligase.

This modified RNA preparation is used as a template for first strand cDNA synthesis using a gene specific oligonucleotide. The first strand synthesis reaction is used as a template for PCR amplification of the desired 5′ end using a primer specific to the ligated RNA oligonucleotide and a primer specific to the known sequence of the gene of interest. The resultant product is then sequenced and analyzed to confirm that the 5′ end sequence belongs to the desired gene.

Example 2 Isolation of Genomic Clones Corresponding to a Polynucleotide

A human genomic P1 library (Genomic Systems, Inc.) is screened by PCR using primers selected for the sequence corresponding to SEQ ID NO:X according to the method described in Example 1. (See also, Sambrook.)

Example 3 Tissue Specific Expression Analysis

The Human Genome Sciences, Inc. (HGS) database is derived from sequencing tissue and/or disease specific cDNA libraries. Libraries generated from a particular tissue are selected and the specific tissue expression pattern of EST groups or assembled contigs within these libraries is determined by comparison of the expression patterns of those groups or contigs within the entire database. ESTs and assembled contigs which show tissue specific expression are selected.

The original clone from which the specific EST sequence was generated, or in the case of an assembled contig, the clone from which the 5′ most EST sequence was generated, is obtained from the catalogued library of clones and the insert amplified by PCR using methods known in the art. The PCR product is denatured and then transferred in 96 or 384 well format to a nylon membrane (Schleicher and Scheull) generating an array filter of tissue specific clones. Housekeeping genes, maize genes, and known tissue specific genes are included on the filters. These targets can be used in signal normalization and to validate assay sensitivity. Additional targets are included to monitor probe length and specificity of hybridization.

Radioactively labeled hybridization probes are generated by first strand cDNA synthesis per the manufacturer's instructions (Life Technologies) from mRNA/RNA samples prepared from the specific tissue being analyzed (e.g., prostate, prostate cancer, ovarian, ovarian cancer, etc.). The hybridization probes are purified by gel exclusion chromatography, quantitated, and hybridized with the array filters in hybridization bottles at 65° C. overnight. The filters are washed under stringent conditions and signals are captured using a Fuji phosphorimager.

Data is extracted using AIS software and following background subtraction, signal normalization is performed. This includes a normalization of filter-wide expression levels between different experimental runs. Genes that are differentially expressed in the tissue of interest are identified.

Example 4 Chromosomal Mapping of the Polynucleotides

An oligonucleotide primer set is designed according to the sequence at the 5′ end of SEQ ID NO:X. This primer preferably spans about 100 nucleotides. This primer set is then used in a polymerase chain reaction under the following set of conditions: 30 seconds, 95° C.; 1 minute, 56° C.; 1 minute, 70° C. This cycle is repeated 32 times followed by one 5 minute cycle at 70° C. Human, mouse, and hamster DNA is used as template in addition to a somatic cell hybrid panel containing individual chromosomes or chromosome fragments (Bios, Inc). The reactions are analyzed on either 8% polyacrylamide gels or 3.5% agarose gels. Chromosome mapping is determined by the presence of an approximately 100 bp PCR fragment in the particular somatic cell hybrid.

Example 5 Bacterial Expression of a Polypeptide

A polynucleotide encoding a polypeptide of the present invention is amplified using PCR oligonucleotide primers corresponding to the 5′ and 3′ ends of the DNA sequence, as outlined in Example 1, to synthesize insertion fragments. The primers used to amplify the cDNA insert should preferably contain restriction sites, such as BamHI and XbaI, at the 5′ end of the primers in order to clone the amplified product into the expression vector. For example, BamHI and XbaI correspond to the restriction enzyme sites on the bacterial expression vector pQE-9. (Qiagen, Inc., Chatsworth, Calif.). This plasmid vector encodes antibiotic resistance (Amp^(r)), a bacterial origin of replication (ori), an IPTG-regulatable promoter/operator (P/O), a ribosome binding site (RBS), a 6-histidine tag (6-His), and restriction enzyme cloning sites.

The pQE-9 vector is digested with BamHI and XbaI and the amplified fragment is ligated into the pQE-9 vector maintaining the reading frame initiated at the bacterial RBS. The ligation mixture is then used to transform the E. coli strain M15/rep4 (Qiagen, Inc.) which contains multiple copies of the plasmid pREP4, which expresses the lacI repressor and also confers kanamycin resistance (Kan^(r)). Transformants are identified by their ability to grow on LB plates and ampicillin/kanamycin resistant colonies are selected. Plasmid DNA is isolated and confirmed by restriction analysis.

Clones containing the desired constructs are grown overnight (O/N) in liquid culture in LB media supplemented with both Amp (100 ug/ml) and Kan (25 ug/ml). The O/N culture is used to inoculate a large culture at a ratio of 1:100 to 1:250. The cells are grown to an optical density 600 (O.D.⁶⁰⁰) of between 0.4 and 0.6. IPTG (Isopropyl-B-D-thiogalacto pyranoside) is then added to a final concentration of 1 mM. IPTG induces by inactivating the lacI repressor, clearing the P/0 leading to increased gene expression.

Cells are grown for an extra 3 to 4 hours. Cells are then harvested by centrifugation (20 mins at 6000×g). The cell pellet is solubilized in the chaotropic agent 6 Molar Guanidine HCl by stirring for 3-4 hours at 4° C. The cell debris is removed by centrifugation, and the supernatant containing the polypeptide is loaded onto a nickel-nitrilo-tri-acetic acid (“Ni-NTA”) affinity resin column (available from QIAGEN, Inc., supra). Proteins with a 6×His tag bind to the Ni-NTA resin with high affinity and can be purified in a simple one-step procedure (for details see: The QIAexpressionist (1995) QIAGEN, Inc., supra).

Briefly, the supernatant is loaded onto the column in 6 M guanidine-HCl, pH 8. The column is first washed with 10 volumes of 6 M guanidine-HCl, pH 8, then washed with 10 volumes of 6 M guanidine-HCl pH 6, and finally the polypeptide is eluted with 6 M guanidine-HCl, pH 5.

The purified protein is then renatured by dialyzing it against phosphate-buffered saline (PBS) or 50 mM Na-acetate, pH 6 buffer plus 200 mM NaCl. Alternatively, the protein can be successfully refolded while immobilized on the Ni-NTA column. The recommended conditions are as follows: renature using a linear 6M-1M urea gradient in 500 mM NaCl, 20% glycerol, 20 mM Tris/HCl pH 7.4, containing protease inhibitors. The renaturation should be performed over a period of 1.5 hours or more. After renaturation the proteins are eluted by the addition of 250 mM immidazole. Immidazole is removed by a final dialyzing step against PBS or 50 mM sodium acetate pH 6 buffer plus 200 mM NaCl. The purified protein is stored at 4° C. or frozen at −80° C.

In addition to the above expression vector, the present invention further includes an expression vector, called pHE4a (ATCC Accession Number 209645, deposited on Feb. 25, 1998) which contains phage operator and promoter elements operatively linked to a polynucleotide of the present invention, called pHE4a. (ATCC Accession Number 209645, deposited on Feb. 25, 1998.) This vector contains: 1) a neomycinphosphotransferase gene as a selection marker, 2) an E. coli origin of replication, 3) a T5 phage promoter sequence, 4) two lac operator sequences, 5) a Shine-Delgarno sequence, and 6) the lactose operon repressor gene (lacIq). The origin of replication (oriC) is derived from pUC19 (LTI, Gaithersburg, Md.). The promoter and operator sequences are made synthetically.

DNA can be inserted into the pHE4a by restricting the vector with NdeI and XbaI, BamHI, XhoI, or Asp718, running the restricted product on a gel, and isolating the larger fragment (the stuffer fragment should be about 310 base pairs). The DNA insert is generated according to the PCR protocol described in Example 1, using PCR primers having restriction sites for NdeI (5′ primer) and XbaI, BamHI, XhoI, or Asp718 (3′ primer). The PCR insert is gel purified and restricted with compatible enzymes. The insert and vector are ligated according to standard protocols.

The engineered vector could easily be substituted in the above protocol to express protein in a bacterial system.

Example 6 Purification of a Polypeptide from an Inclusion Body

The following alternative method can be used to purify a polypeptide expressed in E coli when it is present in the form of inclusion bodies. Unless otherwise specified, all of the following steps are conducted at 4-10° C.

Upon completion of the production phase of the E. coli fermentation, the cell culture is cooled to 4-10° C. and the cells harvested by continuous centrifugation at 15,000 rpm (Heraeus Sepatech). On the basis of the expected yield of protein per unit weight of cell paste and the amount of purified protein required, an appropriate amount of cell paste, by weight, is suspended in a buffer solution containing 100 mM Tris, 50 mM EDTA, pH 7.4. The cells are dispersed to a homogeneous suspension using a high shear mixer.

The cells are then lysed by passing the solution through a microfluidizer (Microfuidics, Corp. or APV Gaulin, Inc.) twice at 4000-6000 psi. The homogenate is then mixed with NaCl solution to a final concentration of 0.5 M NaCl, followed by centrifugation at 7000×g for 15 min. The resultant pellet is washed again using 0.5M NaCl, 100 mM Tris, 50 mM EDTA, pH 7.4.

The resulting washed inclusion bodies are solubilized with 1.5 M guanidine hydrochloride (GuHCl) for 2-4 hours. After 7000×g centrifugation for 15 min., the pellet is discarded and the polypeptide containing supernatant is incubated at 4° C. overnight to allow further GuHCl extraction.

Following high speed centrifugation (30,000×g) to remove insoluble particles, the GuHCl solubilized protein is refolded by quickly mixing the GuHCl extract with 20 volumes of buffer containing 50 mM sodium, pH 4.5, 150 mM NaCl, 2 mM EDTA by vigorous stirring. The refolded diluted protein solution is kept at 4° C. without mixing for 12 hours prior to further purification steps.

To clarify the refolded polypeptide solution, a previously prepared tangential filtration unit equipped with 0.16 μm membrane filter with appropriate surface area (e.g., Filtron), equilibrated with 40 mM sodium acetate, pH 6.0 is employed. The filtered sample is loaded onto a cation exchange resin (e.g., Poros HS-50, Perseptive Biosystems). The column is washed with 40 mM sodium acetate, pH 6.0 and eluted with 250 mM, 500 mM, 1000 mM, and 1500 mM NaCl in the same buffer, in a stepwise manner. The absorbance at 280 nm of the effluent is continuously monitored. Fractions are collected and further analyzed by SDS-PAGE.

Fractions containing the polypeptide are then pooled and mixed with 4 volumes of water. The diluted sample is then loaded onto a previously prepared set of tandem columns of strong anion (Poros HQ-50, Perseptive Biosystems) and weak anion (Poros CM-20, Perseptive Biosystems) exchange resins. The columns are equilibrated with 40 mM sodium acetate, pH 6.0. Both columns are washed with 40 mM sodium acetate, pH 6.0, 200 mM NaCl. The CM-20 column is then eluted using a 10 column volume linear gradient ranging from 0.2 M NaCl, 50 mM sodium acetate, pH 6.0 to 1.0 M NaCl, 50 mM sodium acetate, pH 6.5. Fractions are collected under constant A₂₈₀ monitoring of the effluent. Fractions containing the polypeptide (determined, for instance, by 16% SDS-PAGE) are then pooled.

The resultant polypeptide should exhibit greater than 95% purity after the above refolding and purification steps. No major contaminant bands should be observed from Commassie blue stained 16% SDS-PAGE gel when 5 μg of purified protein is loaded. The purified protein can also be tested for endotoxin/LPS contamination, and typically the LPS content is less than 0.1 ng/ml according to LAL assays.

Example 7 Cloning and Expression of a Polypeptide in a Baculovirus Expression System

In this example, the plasmid shuttle vector pA2 is used to insert a polynucleotide into a baculovirus to express a polypeptide. This expression vector contains the strong polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus (AcMNPV) followed by convenient restriction sites such as BamHI, Xba I and Asp718. The polyadenylation site of the simian virus 40 (“SV40”) is used for efficient polyadenylation. For easy selection of recombinant virus, the plasmid contains the beta-galactosidase gene from E. coli under control of a weak Drosophila promoter in the same orientation, followed by the polyadenylation signal of the polyhedrin gene. The inserted genes are flanked on both sides by viral sequences for cell-mediated homologous recombination with wild-type viral DNA to generate a viable virus that express the cloned polynucleotide.

Many other baculovirus vectors can be used in place of the vector above, such as pAc373, pVL941, and pAcIM1, as one skilled in the art would readily appreciate, as long as the construct provides appropriately located signals for transcription, translation, secretion and the like, including a signal peptide and an in-frame AUG as required. Such vectors are described, for instance, in Luckow et al., Virology 170:31-39 (1989).

Specifically, the cDNA sequence contained in the deposited clone, including the AUG initiation codon, is amplified using the PCR protocol described in Example 1. If a naturally occurring signal sequence is used to produce the polypeptide of the present invention, the pA2 vector does not need a second signal peptide. Alternatively, the vector can be modified (pA2 GP) to include a baculovirus leader sequence, using the standard methods described in Summers et al., “A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures,” Texas Agricultural Experimental Station Bulletin No. 1555 (1987).

The amplified fragment is isolated from a 1% agarose gel using a commercially available kit (“Geneclean,” BIO 101 Inc., La Jolla, Calif.). The fragment then is digested with appropriate restriction enzymes and again purified on a 1% agarose gel.

The plasmid is digested with the corresponding restriction enzymes and optionally, can be dephosphorylated using calf intestinal phosphatase, using routine procedures known in the art. The DNA is then isolated from a 1% agarose gel using a commercially available kit (“Geneclean” BIO 101 Inc., La Jolla, Calif.).

The fragment and the dephosphorylated plasmid are ligated together with T4 DNA ligase. E. coli HB101 or other suitable E. coli hosts such as XL-1 Blue (Stratagene Cloning Systems, La Jolla, Calif.) cells are transformed with the ligation mixture and spread on culture plates. Bacteria containing the plasmid are identified by digesting DNA from individual colonies and analyzing the digestion product by gel electrophoresis. The sequence of the cloned fragment is confirmed by DNA sequencing.

Five μg of a plasmid containing the polynucleotide is co-transfected with 1.0 μg of a commercially available linearized baculovirus DNA (“BaculoGold™ baculovirus DNA, Pharmingen, San Diego, Calif.), using the lipofection method described by Felgner et al., Proc. Natl. Acad. Sci. USA 84:7413-7417 (1987). One μg of BaculoGold™ virus DNA and 5 μg of the plasmid are mixed in a sterile well of a microtiter plate containing 50 μl of serum-free Grace's medium (Life Technologies Inc., Gaithersburg, Md.). Afterwards, 10 μl Lipofectin plus 90 μl Grace's medium are added, mixed and incubated for 15 minutes at room temperature. Then the transfection mixture is added drop-wise to Sf9 insect cells (ATCC CRL 1711) seeded in a 35 mm tissue culture plate with 1 ml Grace's medium without serum. The plate is then incubated for 5 hours at 27° C. The transfection solution is then removed from the plate and 1 ml of Grace's insect medium supplemented with 10% fetal calf serum is added. Cultivation is then continued at 27° C. for four days.

After four days the supernatant is collected and a plaque assay is performed, as described by Summers and Smith, supra. An agarose gel with “Blue Gal” (Life Technologies Inc., Gaithersburg) is used to allow easy identification and isolation of gal-expressing clones, which produce blue-stained plaques. (A detailed description of a “plaque assay” of this type can also be found in the user's guide for insect cell culture and baculovirology distributed by Life Technologies Inc., Gaithersburg, page 9-10.) After appropriate incubation, blue stained plaques are picked with the tip of a micropipettor (e.g., Eppendorf). The agar containing the recombinant viruses is then resuspended in a microcentrifuge tube containing 200 μl of Grace's medium and the suspension containing the recombinant baculovirus is used to infect Sf9 cells seeded in 35 mm dishes. Four days later the supernatants of these culture dishes are harvested and then they are stored at 4° C.

To verify the expression of the polypeptide, Sf9 cells are grown in Grace's medium supplemented with 10% heat-inactivated FBS. The cells are infected with the recombinant baculovirus containing the polynucleotide at a multiplicity of infection (“MOI”) of about 2. If radiolabeled proteins are desired, 6 hours later the medium is removed and is replaced with SF900 II medium minus methionine and cysteine (available from Life Technologies Inc., Rockville, Md.). After 42 hours, 5 μCi of ³⁵S-methionine and 5 μCi ³⁵S-cysteine (available from Amersham) are added. The cells are further incubated for 16 hours and then are harvested by centrifugation. The proteins in the supernatant as well as the intracellular proteins are analyzed by SDS-PAGE followed by autoradiography (if radiolabeled).

Microsequencing of the amino acid sequence of the amino terminus of purified protein may be used to determine the amino terminal sequence of the produced protein.

Example 8 Expression of a Polypeptide in Mammalian Cells

The polypeptide of the present invention can be expressed in a mammalian cell. A typical mammalian expression vector contains a promoter element, which mediates the initiation of transcription of mRNA, a protein coding sequence, and signals required for the termination of transcription and polyadenylation of the transcript. Additional elements include enhancers, Kozak sequences and intervening sequences flanked by donor and acceptor sites for RNA splicing. Highly efficient transcription is achieved with the early and late promoters from SV40, the long terminal repeats (LTRs) from Retroviruses, e.g., RSV, HTLVI, HIVI and the early promoter of the cytomegalovirus (CMV). However, cellular elements can also be used (e.g., the human actin promoter).

Suitable expression vectors for use in practicing the present invention include, for example, vectors such as pSVL and pMSG (Pharmacia, Uppsala, Sweden), pRSVcat (ATCC 37152), pSV2dhfr (ATCC 37146), pBC12MI (ATCC 67109), pCMVSport 2.0, and pCMVSport 3.0. Mammalian host cells that could be used include, human Hela, 293, H9 and Jurkat cells, mouse NIH3T3 and C127 cells, Cos 1, Cos 7 and CVI, quail QC1-3 cells, mouse L cells and Chinese hamster ovary (CHO) cells.

Alternatively, the polypeptide can be expressed in stable cell lines containing the polynucleotide integrated into a chromosome. The co-transfection with a selectable marker such as DHFR, gpt, neomycin, or hygromycin allows the identification and isolation of the transfected cells.

The transfected gene can also be amplified to express large amounts of the encoded protein. The DHFR (dihydrofolate reductase) marker is useful in developing cell lines that carry several hundred or even several thousand copies of the gene of interest. (See, e.g., Alt, F. W., et al., J. Biol. Chem. 253:1357-1370 (1978); Hamlin, J. L. and Ma, C., Biochem. et Biophys. Acta, 1097:107-143 (1990); Page, M. J. and Sydenham, M. A., Biotechnology 9:64-68 (1991)). Another useful selection marker is the enzyme glutamine synthase (GS) (Murphy et al., Biochem J. 227:277-279 (1991); Bebbington et al., Bio/Technology 10:169-175 (1992). Using these markers, the mammalian cells are grown in selective medium and the cells with the highest resistance are selected. These cell lines contain the amplified gene(s) integrated into a chromosome. Chinese hamster ovary (CHO) and NSO cells are often used for the production of proteins.

Derivatives of the plasmid pSV2-dhfr (ATCC Accession No. 37146), the expression vectors pC4 (ATCC Accession No. 209646) and pC6 (ATCC Accession No. 209647) contain the strong promoter (LTR) of the Rous Sarcoma Virus (Cullen et al., Molecular and Cellular Biology, 438-447 (March, 1985)) plus a fragment of the CMV-enhancer (Boshart et al., Cell 41:521-530 (1985)). Multiple cloning sites, e.g., with the restriction enzyme cleavage sites BamHI, XbaI and Asp718, facilitate the cloning of the gene of interest. The vectors also contain the 3′ intron, the polyadenylation and termination signal of the rat preproinsulin gene, and the mouse DHFR gene under control of the SV40 early promoter.

Specifically, the plasmid pC6, for example, is digested with appropriate restriction enzymes and then dephosphorylated using calf intestinal phosphates by procedures known in the art. The vector is then isolated from a 1% agarose gel.

A polynucleotide of the present invention is amplified according to the protocol outlined in Example 1. If a naturally occurring signal sequence is used to produce the polypeptide of the present invention, the vector does not need a second signal peptide. Alternatively, if a naturally occurring signal sequence is not used, the vector can be modified to include a heterologous signal sequence. (See, e.g., International Publication No. WO 96/34891.)

The amplified fragment is isolated from a 1% agarose gel using a commercially available kit (“Geneclean,” BIO 101 Inc., La Jolla, Calif.). The fragment then is digested with appropriate restriction enzymes and again purified on a 1% agarose gel.

The amplified fragment is then digested with the same restriction enzyme and purified on a 1% agarose gel. The isolated fragment and the dephosphorylated vector are then ligated with T4 DNA ligase. E. coli HB101 or XL-1 Blue cells are then transformed and bacteria are identified that contain the fragment inserted into plasmid pC6 using, for instance, restriction enzyme analysis.

Chinese hamster ovary cells lacking an active DHFR gene is used for transfection. Five μg of the expression plasmid pC6 or pC4 is cotransfected with 0.5 μg of the plasmid pSVneo using lipofectin (Felgner et al., supra). The plasmid pSV2-neo contains a dominant selectable marker, the neo gene from Tn5 encoding an enzyme that confers resistance to a group of antibiotics including G418. The cells are seeded in alpha minus MEM supplemented with 1 mg/ml G418. After 2 days, the cells are trypsinized and seeded in hybridoma cloning plates (Greiner, Germany) in alpha minus MEM supplemented with 10, 25, or 50 ng/ml of methotrexate plus 1 mg/ml G418. After about 10⁻¹⁴ days single clones are trypsinized and then seeded in 6-well petri dishes or 10 ml flasks using different concentrations of methotrexate (50 nM, 100 nM, 200 nM, 400 nM, 800 nM). Clones growing at the highest concentrations of methotrexate are then transferred to new 6-well plates containing even higher concentrations of methotrexate (1 μM, 2 μM, 5 μM, 10 mM, 20 mM). The same procedure is repeated until clones are obtained which grow at a concentration of 100-200 μM. Expression of the desired gene product is analyzed, for instance, by SDS-PAGE and Western blot or by reversed phase HPLC analysis.

Example 9 Protein Fusions

The polypeptides of the present invention are preferably fused to other proteins. These fusion proteins can be used for a variety of applications. For example, fusion of the present polypeptides to His-tag, HA-tag, protein A, IgG domains, and maltose binding protein facilitates purification. (See Example 5; see also EP A 394,827; Traunecker, et al., Nature 331:84-86 (1988)). Similarly, fusion to IgG-1, IgG-3, and albumin increases the halflife time in vivo. Nuclear localization signals fused to the polypeptides of the present invention can target the protein to a specific subcellular localization, while covalent heterodimer or homodimers can increase or decrease the activity of a fusion protein. Fusion proteins can also create chimeric molecules having more than one function. Finally, fusion proteins can increase solubility and/or stability of the fused protein compared to the non-fused protein. All of the types of fusion proteins described above can be made by modifying the following protocol, which outlines the fusion of a polypeptide to an IgG molecule, or the protocol described in Example 5.

Briefly, the human Fc portion of the IgG molecule can be PCR amplified, using primers that span the 5′ and 3′ ends of the sequence described below. These primers also should have convenient restriction enzyme sites that will facilitate cloning into an expression vector, preferably a mammalian expression vector.

For example, if pC4 (ATCC Accession No. 209646) is used, the human Fc portion can be ligated into the BamHI cloning site. Note that the 3′ BamHI site should be destroyed. Next, the vector containing the human Fc portion is re-restricted with BamHI, linearizing the vector, and a polynucleotide of the present invention, isolated by the PCR protocol described in Example 1, is ligated into this BamHI site. Note that the polynucleotide is cloned without a stop codon, otherwise a fusion protein will not be produced.

If the naturally occurring signal sequence is used to produce the polypeptide of the present invention, pC4 does not need a second signal peptide. Alternatively, if the naturally occurring signal sequence is not used, the vector can be modified to include a heterologous signal sequence. (See, e.g., International Publication No. WO 96/34891.) Human IgG Fc region: (SEQ ID NO: 1) GGGATCCGGAGCCCAAATCTTCTGACAAAACTCACACATGCCCACCGTGC CCAGCACCTGAATTCGAGGGTGCACCGTCAGTCTTCCTCTTCCCCCCAAA ACCCAAGGACACCCTCATGATCTCCCGGACTCCTGAGGTCACATGCGTGG TGGTGGACGTAAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTG GACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTA CAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACT GGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCA ACCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGG TCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCAAGCGACATCGCCGTG GAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCC CGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGG TAAATGAGTGCGACGGCCGCGACTCTAGAGGAT

Example 10 Production of an Antibody from a Polypeptide

a) Hybridoma Technology

The antibodies of the present invention can be prepared by a variety of methods. (See, Current Protocols, Chapter 2.) As one example of such methods, cells expressing a polypeptide of the present invention are administered to an animal to induce the production of sera containing polyclonal antibodies. In a preferred method, a preparation of a polypeptide of the present invention is prepared and purified to render it substantially free of natural contaminants. Such a preparation is then introduced into an animal in order to produce polyclonal antisera of greater specific activity.

Monoclonal antibodies specific for a polypeptide of the present invention are prepared using hybridoma technology (Kohler et al., Nature 256:495 (1975); Kohler et al., Eur. J. Immunol. 6:511 (1976); Kohler et al., Eur. J. Immunol. 6:292 (1976); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas, Elsevier, N.Y., pp. 563-681 (1981)). In general, an animal (preferably a mouse) is immunized with a polypeptide of the present invention or, more preferably, with a secreted polypeptide-expressing cell. Such polypeptide-expressing cells are cultured in any suitable tissue culture medium, preferably in Earle's modified Eagle's medium supplemented with 10% fetal bovine serum (inactivated at about 56° C.), and supplemented with about 10 g/l of nonessential amino acids, about 1,000 U/ml of penicillin, and about 100 μg/ml of streptomycin.

The splenocytes of such mice are extracted and fused with a suitable myeloma cell line. Any suitable myeloma cell line may be employed in accordance with the present invention; however, it is preferable to employ the parent mycloma cell line (SP2O), available from the ATCC. After fusion, the resulting hybridoma cells are selectively maintained in HAT medium, and then cloned by limiting dilution as described by Wands et al. (Gastroenterology 80:225-232 (1981)). The hybridoma cells obtained through such a selection are then assayed to identify clones which secrete antibodies capable of binding the polypeptide of the present invention.

Alternatively, additional antibodies capable of binding to a polypeptide of the present invention can be produced in a two-step procedure using anti-idiotypic antibodies. Such a method makes use of the fact that antibodies are themselves antigens, and therefore, it is possible to obtain an antibody which binds to a second antibody. In accordance with this method, protein specific antibodies are used to immunize an animal, preferably a mouse. The splenocytes of such an animal are then used to produce hybridoma cells, and the hybridoma cells are screened to identify clones which produce an antibody whose ability to bind to the polypeptide-specific antibody can be blocked by said polypeptide. Such antibodies comprise anti-idiotypic antibodies to the polypeptide-specific antibody and are used to immunize an animal to induce formation of further polypeptide-specific antibodies.

For in vivo use of antibodies in humans, an antibody is “humanized”. Such antibodies can be produced using genetic constructs derived from hybridoma cells producing the monoclonal antibodies described above. Methods for producing chimeric and humanized antibodies are known in the art and are discussed herein. (See, for review, Morrison, Science 229:1202 (1985); Oi et al., BioTechniques 4:214 (1986); Cabilly et al., U.S. Pat. No. 4,816,567; Taniguchi et al., EP 171496; Morrison et al., EP 173494; Neuberger et al., WO 8601533; Robinson et al., International Publication No. WO 8702671; Boulianne et al., Nature 312:643 (1984); Neuberger et al., Nature 314:268 (1985)).

b) Isolation of Antibody Fragments Directed Against a Polypeptide of the Present Invention from a Library of scFvs

Naturally occurring V-genes isolated from human PBLs are constructed into a library of antibody fragments which contain reactivities against a polypeptide of the present invention to which the donor may or may not have been exposed (see e.g., U.S. Pat. No. 5,885,793 incorporated herein by reference in its entirety).

Rescue of the Library. A library of scFvs is constructed from the RNA of human PBLs as described in International Publication No. WO 92/01047. To rescue phage displaying antibody fragments, approximately 10⁹ E. coli harboring the phagemid are used to inoculate 50 ml of 2×TY containing 1% glucose and 100 μg/ml of ampicillin (2×TY-AMP-GLU) and grown to an O.D. of 0.8 with shaking. Five ml of this culture is used to inoculate 50 ml of 2×TY-AMP-GLU, 2×108 TU of delta gene 3 helper (M13 delta gene III, see International Publication No. WO 92/01047) are added and the culture incubated at 37° C. for 45 minutes without shaking and then at 37° C. for 45 minutes with shaking. The culture is centrifuged at 4000 r.p.m. for 10 min. and the pellet resuspended in 2 liters of 2×TY containing 100 μg/ml ampicillin and 50 ug/ml kanamycin and grown overnight. Phage are prepared as described in International Publication No. WO 92/01047.

M13 delta gene III is prepared as follows: M13 delta gene III helper phage does not encode gene III protein, hence the phage(mid) displaying antibody fragments have a greater avidity of binding to antigen. Infectious M13 delta gene III particles are made by growing the helper phage in cells harboring a pUC19 derivative supplying the wild type gene III protein during phage morphogenesis. The culture is incubated for 1 hour at 37° C. without shaking and then for a further hour at 37° C. with shaking. Cells are spun down (IEC-Centra 8,400 r.p.m. for 10 min), resuspended in 300 ml 2×TY broth containing 100 μg ampicillin/ml and 25 μg kanamycin/ml (2×TY-AMP-KAN) and grown overnight, shaking at 37° C. Phage particles are purified and concentrated from the culture medium by two PEG-precipitations (Sambrook et al., 1990), resuspended in 2 ml PBS and passed through a 0.45 μm filter (Minisart NML; Sartorius) to give a final concentration of approximately 1013 transducing units/ml (ampicillin-resistant clones).

Panning of the Library. Immunotubes (Nunc) are coated overnight in PBS with 4 ml of either 100 μg/ml or 10 μg/ml of a polypeptide of the present invention. Tubes are blocked with 2% Marvel-PBS for 2 hours at 37° C. and then washed 3 times in PBS. Approximately 10¹³ TU of phage is applied to the tube and incubated for 30 minutes at room temperature tumbling on an over and under turntable and then left to stand for another 1.5 hours. Tubes are washed 10 times with PBS 0.1% Tween-20 and 10 times with PBS. Phage are eluted by adding 1 ml of 100 mM triethylamine and rotating 15 minutes on an under and over turntable after which the solution is immediately neutralized with 0.5 ml of 1.0M Tris-HCl, pH 7.4. Phage are then used to infect 10 ml of mid-log E. coli TG1 by incubating eluted phage with bacteria for 30 minutes at 37° C. The E. coli are then plated on TYE plates containing 1% glucose and 100 μg/ml ampicillin. The resulting bacterial library is then rescued with delta gene 3 helper phage as described above to prepare phage for a subsequent round of selection. This process is then repeated for a total of 4 rounds of affinity purification with tube-washing increased to 20 times with PBS, 0.1% Tween-20 and 20 times with PBS for rounds 3 and 4.

Characterization of Binders. Eluted phage from the 3rd and 4th rounds of selection are used to infect E. coli HB 2151 and soluble scFv is produced (Marks, et al., 1991) from single colonies for assay. ELISAs are performed with microtitre plates coated with either 10 pg/ml of the polypeptide of the present invention in 50 mM bicarbonate pH 9.6. Clones positive in ELISA are further characterized by PCR fingerprinting (see, e.g., International Publication No. WO 92/01047) and then by sequencing. These ELISA positive clones may also be further characterized by techniques known in the art, such as, for example, epitope mapping, binding affinity, receptor signal transduction, ability to block or competitively inhibit antibody/antigen binding, and competitive agonistic or antagonistic activity.

Example 11 Method of Determining Alterations in a Gene Corresponding to a Polynucleotide

RNA isolated from entire families or individual patients presenting with a phenotype of interest (such as a disease) is isolated. cDNA is then generated from these RNA samples using protocols known in the art. (See, Sambrook.) The cDNA is then used as a template for PCR, employing primers surrounding regions of interest in SEQ ID NO:X; and/or the nucleotide sequence of the cDNA contained in ATCC Deposit No:Z. Suggested PCR conditions consist of 35 cycles at 95 degrees C. for 30 seconds; 60-120 seconds at 52-58 degrees C.; and 60-120 seconds at 70 degrees C., using buffer solutions described in Sidransky et al., Science 252:706 (1991).

PCR products are then sequenced using primers labeled at their 5′ end with T4 polynucleotide kinase, employing SequiTherm Polymerase (Epicentre Technologies). The intron-exon boundaries of selected exons is also determined and genomic PCR products analyzed to confirm the results. PCR products harboring suspected mutations are then cloned and sequenced to validate the results of the direct sequencing.

PCR products are cloned into T-tailed vectors as described in Holton et al., Nucleic Acids Research, 19:1156 (1991) and sequenced with T7 polymerase (United States Biochemical). Affected individuals are identified by mutations not present in unaffected individuals.

Genomic rearrangements are also observed as a method of determining alterations in a gene corresponding to a polynucleotide. Genomic clones isolated according to Example 2 are nick-translated with digoxigenindeoxy-uridine 5′-triphosphate (Boehringer Manheim), and FISH performed as described in Johnson et al., Methods Cell Biol. 35:73-99 (1991). Hybridization with the labeled probe is carried out using a vast excess of human cot-1 DNA for specific hybridization to the corresponding genomic locus.

Chromosomes are counterstained with 4,6-diamino-2-phenylidole and propidium iodide, producing a combination of C- and R-bands. Aligned images for precise mapping are obtained using a triple-band filter set (Chroma Technology, Brattleboro, Vt.) in combination with a cooled charge-coupled device camera (Photometrics, Tucson, Ariz.) and variable excitation wavelength filters. (Johnson et al., Genet. Anal. Tech. Appl., 8:75 (1991)). Image collection, analysis and chromosomal fractional length measurements are performed using the ISee Graphical Program System. (Inovision Corporation, Durham, N.C.) Chromosome alterations of the genomic region hybridized by the probe are identified as insertions, deletions, and translocations. These alterations are used as a diagnostic marker for an associated disease.

Example 12 Method of Detecting Abnormal Levels of a Polypeptide in a Biological Sample

A polypeptide of the present invention can be detected in a biological sample, and if an increased or decreased level of the polypeptide is detected, this polypeptide is a marker for a particular phenotype. Methods of detection are numerous, and thus, it is understood that one skilled in the art can modify the following assay to fit their particular needs.

For example, antibody-sandwich ELISAs are used to detect polypeptides in a sample, preferably a biological sample. Wells of a microtiter plate are coated with specific antibodies, at a final concentration of 0.2 to 10 ug/ml. The antibodies are either monoclonal or polyclonal and are produced by the method described in Example 10. The wells are blocked so that non-specific binding of the polypeptide to the well is reduced.

The coated wells are then incubated for >2 hours at RT with a sample containing the polypeptide. Preferably, serial dilutions of the sample should be used to validate results. The plates are then washed three times with deionized or distilled water to remove unbound polypeptide.

Next, 50 ul of specific antibody-alkaline phosphatase conjugate, at a concentration of 25-400 ng, is added and incubated for 2 hours at room temperature. The plates are again washed three times with deionized or distilled water to remove unbound conjugate.

Add 75 ul of 4-methylumbelliferyl phosphate (MUP) or p-nitrophenyl phosphate (NPP) substrate solution to each well and incubate 1 hour at room temperature. Measure the reaction by a microtiter plate reader. Prepare a standard curve, using serial dilutions of a control sample, and plot polypeptide concentration on the X-axis (log scale) and fluorescence or absorbance of the Y-axis (linear scale). Interpolate the concentration of the polypeptide in the sample using the standard curve.

Example 13 Formulation

The invention also provides methods of treatment and/or prevention of diseases or disorders (such as, for example, any one or more of the diseases or disorders disclosed herein) by administration to a subject of an effective amount of a Therapeutic. By therapeutic is meant polynucleotides or polypeptides of the invention (including fragments and variants), agonists or antagonists thereof, and/or antibodies thereto, in combination with a pharmaceutically acceptable carrier type (e.g., a sterile carrier).

The Therapeutic will be formulated and dosed in a fashion consistent with good medical practice, taking into account the clinical condition of the individual patient (especially the side effects of treatment with the Therapeutic alone), the site of delivery, the method of administration, the scheduling of administration, and other factors known to practitioners. The “effective amount” for purposes herein is thus determined by such considerations.

As a general proposition, the total pharmaceutically effective amount of the Therapeutic administered parenterally per dose will be in the range of about 1 ug/kg/day to 10 mg/kg/day of patient body weight, although, as noted above, this will be subject to therapeutic discretion. More preferably, this dose is at least 0.01 mg/kg/day, and most preferably for humans between about 0.01 and 1 mg/kg/day for the hormone. If given continuously, the Therapeutic is typically administered at a dose rate of about 1 ug/kg/hour to about 50 ug/kg/hour, either by 1-4 injections per day or by continuous subcutaneous infusions, for example, using a mini-pump. An intravenous bag solution may also be employed. The length of treatment needed to observe changes and the interval following treatment for responses to occur appears to vary depending on the desired effect.

Therapeutics can be are administered orally, rectally, parenterally, intracistemally, intravaginally, intraperitoneally, topically (as by powders, ointments, gels, drops or transdermal patch), bucally, or as an oral or nasal spray. “Pharmaceutically acceptable carrier” refers to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any. The term “parenteral” as used herein refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.

Therapeutics of the invention are also suitably administered by sustained-release systems. Suitable examples of sustained-release Therapeutics are administered orally, rectally, parenterally, intracistemally, intravaginally, intraperitoneally, topically (as by powders, ointments, gels, drops or transdermal patch), bucally, or as an oral or nasal spray. “Pharmaceutically acceptable carrier” refers to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. The term “parenteral” as used herein refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.

Therapeutics of the invention are also suitably administered by sustained-release systems. Suitable examples of sustained-release Therapeutics include suitable polymeric materials (such as, for example, semi-permeable polymer matrices in the form of shaped articles, e.g., films, or mirocapsules), suitable hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, and sparingly soluble derivatives (such as, for example, a sparingly soluble salt).

Sustained-release matrices include polylactides (U.S. Pat. No. 3,773,919, EP 58,481), copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman et al., Biopolymers 22:547-556 (1983)), poly (2-hydroxyethyl methacrylate) (Langer et al., J. Biomed. Mater. Res. 15:167-277 (1981), and Langer, Chem. Tech. 12:98-105 (1982)), ethylene vinyl acetate (Langer et al., Id.) or poly-D-(−)-3-hydroxybutyric acid (EP 133,988).

In a preferred embodiment, polypeptide, polynucleotide, and antibody compositions of the invention are formulated in a biodegradable, polymeric drug delivery system, for example as described in U.S. Pat. Nos. 4,938,763; 5,278,201; 5,278,202; 5,324,519; 5,340,849; and 5,487,897 and in International Publication Numbers WO01/35929, WO00/24374, and WO00/06117 which are hereby incorporated by reference in their entirety. In specific preferred embodiments the polypeptide, polynucleotide, and antibody compositions of the invention are formulated using the ATRIGEL® Biodegradable System of Atrix Laboratories, Inc. (Fort Collins, Colo.).

Examples of biodegradable polymers which can be used in the formulation of polypeptide, polynucleotide, and antibody compositions, include but are not limited to, polylactides, polyglycolides, polycaprolactones, polyanhydrides, polyamides, polyurethanes, polyesteramides, polyorthoesters, polydioxanones, polyacetals, polyketals, polycarbonates, polyorthocarbonates, polyphosphazenes, polyhydroxybutyrates, polyhydroxyvalerates, polyalkylene oxalates, polyalkylene succinates, poly(malic acid), poly(amino acids), poly(methyl vinyl ether), poly(maleic anhydride), polyvinylpyrrolidone, polyethylene glycol, polyhydroxycellulose, chitin, chitosan, and copolymers, telpolymers, or combinations or mixtures of the above materials. The preferred polymers are those that have a lower degree of crystallization and are more hydrophobic. These polymers and copolymers are more soluble in the biocompatible solvents than the highly crystalline polymers such as polyglycolide and chitin which also have a high degree of hydrogen-bonding. Preferred materials with the desired solubility parameters are the polylactides, polycaprolactones, and copolymers of these with glycolide in which there are more amorphous regions to enhance solubility. In specific preferred embodiments, the biodegradable polymers which can be used in the formulation of polypeptide, polynucleotide, and antibody compositions are poly(lactide-co-glycolides). Polymer properties such as molecular weight, hydrophobicity, and lactide/glycolide ratio may be modified to obtain the desired polypeptide, polynucleotide, or antibody release profile (See, e.g., Ravivarapu et al., Journal of Pharmaceutical Sciences 89:732-741 (2000), which is hereby incorporated by reference in its entirety).

It is also preferred that the solvent for the biodegradable polymer be non-toxic, water miscible, and otherwise biocompatible. Examples of such solvents include, but are not limited to, N-methyl-2-pyrrolidone, 2-pyrrolidone, C2 to C6 alkanols, C1 to C15 alchohols, dils, triols, and tetraols such as ethanol, glycerine propylene glycol, butanol; C3 to C15 alkyl ketones such as acetone, diethyl ketone and methyl ethyl ketone; C3 to C15 esters such as methyl acetate, ethyl acetate, ethyl lactate; alkyl ketones such as methyl ethyl ketone, C1 to C15 amides such as dimethylformamide, dimethylacetamide and caprolactam; C3 to C20 ethers such as tetrahydrofuran, or solketal; tweens, triacetin, propylene carbonate, decylmethylsulfoxide, dimethyl sulfoxide, oleic acid, 1-dodecylazacycloheptan-2-one, Other preferred solvents are benzyl alchohol, benzyl benzoate, dipropylene glycol, tributyrin, ethyl oleate, glycerin, glycofural, isopropyl myristate, isopropyl palmitate, oleic acid, polyethylene glycol, propylene carbonate, and triethyl citrate. The most preferred solvents are N-methyl-2-pyrrolidone, 2-pyrrolidone, dimethyl sulfoxide, triacetin, and propylene carbonate because of the solvating ability and their compatibility.

Additionally, formulations comprising polypeptide, polynucleotide, and antibody compositions and a biodegradable polymer may also include release-rate modification agents and/or pore-forming agents. Examples of release-rate modification agents include, but are not limited to, fatty acids, triglycerides, other like hydrophobic compounds, organic solvents, plasticizing compounds and hydrophilic compounds. Suitable release rate modification agents include, for example, esters of mono-, di-, and tricarboxylic acids, such as 2-ethoxyethyl acetate, methyl acetate, ethyl acetate, diethyl phthalate, dimethyl phthalate, dibutyl phthalate, dimethyl adipate, dimethyl succinate, dimethyl oxalate, dimethyl citrate, triethyl citrate, acetyl tributyl citrate, acetyl triethyl citrate, glycerol triacetate, di(n-butyl) sebecate, and the like; polyhydroxy alcohols, such as propylene glycol, polyethylene glycol, glycerin, sorbitol, and the like; fatty acids; triesters of glycerol, such as triglycerides, epoxidized soybean oil, and other epoxidized vegetable oils; sterols, such as cholesterol; alcohols, such as C.sub.6-C.sub.12 alkanols, 2-ethoxyethanol. The release rate modification agent may be used singly or in combination with other such agents. Suitable combinations of release rate modification agents include, but are not limited to, glycerin/propylene glycol, sorbitol/glycerine, ethylene oxide/propylene oxide, butylene glycol/adipic acid, and the like. Preferred release rate modification agents include, but are not limited to, dimethyl citrate, triethyl citrate, ethyl heptanoate, glycerin, and hexanediol. Suitable pore-forming agents that may be used in the polymer composition include, but are not limited to, sugars such as sucrose and dextrose, salts such as sodium chloride and sodium carbonate, polymers such as hydroxylpropylcellulose, carboxymethylcellulose, polyethylene glycol, and polyvinylpyrrolidone. Solid crystals that will provide a defined pore size, such as salt or sugar, are preferred.

In specific preferred embodiments the polypeptide, polynucleotide, and antibody compositions of the invention are formulated using the BEMA™ BioErodible Mucoadhesive System, MCA™ MucoCutaneous Absorption System, SMP™ Solvent MicroParticle System, or BCP™ BioCompatible Polymer System of Atrix Laboratories, Inc. (Fort Collins, Colo.).

Sustained-release Therapeutics also include liposomally entrapped Therapeutics of the invention (see generally, Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 317-327 and 353-365 (1989)). Liposomes containing the Therapeutic are prepared by methods known per se: DE 3,218,121; Epstein et al., Proc. Natl. Acad. Sci. (USA) 82:3688-3692 (1985); Hwang et al., Proc. Natl. Acad. Sci. (USA) 77:4030-4034 (1980); EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP 142,641; Japanese Pat. Appl. 83-118008; U.S. Pat. Nos. 4,485,045 and 4,544,545; and EP 102,324. Ordinarily, the liposomes are of the small (about 200-800 Angstroms) unilamellar type in which the lipid content is greater than about 30 mol. percent cholesterol, the selected proportion being adjusted for the optimal Therapeutic.

In yet an additional embodiment, the Therapeutics of the invention are delivered by way of a pump (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989)).

Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990)).

For parenteral administration, in one embodiment, the Therapeutic is formulated generally by mixing it at the desired degree of purity, in a unit dosage injectable form (solution, suspension, or emulsion), with a pharmaceutically acceptable carrier, i.e., one that is non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation. For example, the formulation preferably does not include oxidizing agents and other compounds that are known to be deleterious to the Therapeutic.

Generally, the formulations are prepared by contacting the Therapeutic uniformly and intimately with liquid carriers or finely divided solid carriers or both. Then, if necessary, the product is shaped into the desired formulation. Preferably the carrier is a parenteral carrier, more preferably a solution that is isotonic with the blood of the recipient. Examples of such carrier vehicles include water, saline, Ringer's solution, and dextrose solution. Non-aqueous vehicles such as fixed oils and ethyl oleate are also useful herein, as well as liposomes.

The carrier suitably contains minor amounts of additives such as substances that enhance isotonicity and chemical stability. Such materials are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, succinate, acetic acid, and other organic acids or their salts; antioxidants such as ascorbic acid; low molecular weight (less than about ten residues) polypeptides, e.g., polyarginine or tripeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids, such as glycine, glutamic acid, aspartic acid, or arginine; monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, manose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; counterions such as sodium; and/or nonionic surfactants such as polysorbates, poloxamers, or PEG.

The Therapeutic is typically formulated in such vehicles at a concentration of about 0.1 mg/ml to 100 mg/ml, preferably 1-10 mg/ml, at a pH of about 3 to 8. It will be understood that the use of certain of the foregoing excipients, carriers, or stabilizers will result in the formation of polypeptide salts.

Any pharmaceutical used for therapeutic administration can be sterile. Sterility is readily accomplished by filtration through sterile filtration membranes (e.g., 0.2 micron membranes). Therapeutics generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

Therapeutics ordinarily will be stored in unit or multi-dose containers, for example, sealed ampoules or vials, as an aqueous solution or as a lyophilized formulation for reconstitution. As an example of a lyophilized formulation, 10-ml vials are filled with 5 ml of sterile-filtered 1% (w/v) aqueous Therapeutic solution, and the resulting mixture is lyophilized. The infusion solution is prepared by reconstituting the lyophilized Therapeutic using bacteriostatic Water-for-Injection.

The invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the Therapeutics of the invention. Associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration. In addition, the Therapeutics may be employed in conjunction with other therapeutic compounds.

The Therapeutics of the invention may be administered alone or in combination with adjuvants. Adjuvants that may be administered with the Therapeutics of the invention include, but are not limited to, alum, alum plus deoxycholate (ImmunoAg), MTP-PE (Biocine Corp.), QS21 (Genentech, Inc.), BCG (e.g., THERACYS®), MPL and nonviable prepartions of Corynebacterium parvum. In a specific embodiment, Therapeutics of the invention are administered in combination with alum. In another specific embodiment, Therapeutics of the invention are administered in combination with QS-21. Further adjuvants that may be administered with the Therapeutics of the invention include, but are not limited to, Monophosphoryl lipid immunomodulator, AdjuVax 100a, QS-21, QS-18, CRL1005, Aluminum salts, MF-59, and Virosomal adjuvant technology. Vaccines that may be administered with the Therapeutics of the invention include, but are not limited to, vaccines directed toward protection against MMR (measles, mumps, rubella), polio, varicella, tetanus/diptheria, hepatitis A, hepatitis B, haemophilus influenzae B, whooping cough, pneumonia, influenza, Lyme's Disease, rotavirus, cholera, yellow fever, Japanese encephalitis, poliomyelitis, rabies, typhoid fever, and pertussis. Combinations may be administered either concomitantly, e.g., as an admixture, separately but simultaneously or concurrently; or sequentially. This includes presentations in which the combined agents are administered together as a therapeutic mixture, and also procedures in which the combined agents are administered separately but simultaneously, e.g., as through separate intravenous lines into the same individual. Administration “in combination” further includes the separate administration of one of the compounds or agents given first, followed by the second.

The Therapeutics of the invention may be administered alone or in combination with other therapeutic agents. Therapeutic agents that may be administered in combination with the Therapeutics of the invention, include but not limited to, chemotherapeutic agents, antibiotics, steroidal and non-steroidal anti-inflammatories, conventional immunotherapeutic agents, and/or therapeutic treatments described below. Combinations may be administered either concomitantly, e.g., as an admixture, separately but simultaneously or concurrently; or sequentially. This includes presentations in which the combined agents are administered together as a therapeutic mixture, and also procedures in which the combined agents are administered separately but simultaneously, e.g., as through separate intravenous lines into the same individual. Administration “in combination” further includes the separate administration of one of the compounds or agents given first, followed by the second.

In one embodiment, the Therapeutics of the invention are administered in combination with an anticoagulant. Anticoagulants that may be administered with the compositions of the invention include, but are not limited to, heparin, low molecular weight heparin, warfarin sodium (e.g., COUMADIN®), dicumarol, 4-hydroxycoumarin, anisindione (e.g., MIRADON™), acenocoumarol (e.g., nicoumalone, SINTHROME™), indan-1,3-dione, phenprocoumon (e.g., MARCUMAR™), ethyl biscoumacetate (e.g., TROMEXAN™), and aspirin. In a specific embodiment, compositions of the invention are administered in combination with heparin and/or warfarin. In another specific embodiment, compositions of the invention are administered in combination with warfarin. In another specific embodiment, compositions of the invention are administered in combination with warfarin and aspirin. In another specific embodiment, compositions of the invention are administered in combination with heparin. In another specific embodiment, compositions of the invention are administered in combination with heparin and aspirin.

In another embodiment, the Therapeutics of the invention are administered in combination with thrombolytic drugs. Thrombolytic drugs that may be administered with the compositions of the invention include, but are not limited to, plasminogen, lys-plasminogen, alpha2-antiplasmin, streptokinae (e.g., KABIKINASE™), antiresplace (e.g., EMINASE™), tissue plasminogen activator (t-PA, altevase, ACTIVASE™), urokinase (e.g., ABBOKINASE™), sauruplase, (Prourokinase, single chain urokinase), and aminocaproic acid (e.g., AMICAR™). In a specific embodiment, compositions of the invention are administered in combination with tissue plasminogen activator and aspirin.

In another embodiment, the Therapeutics of the invention are administered in combination with antiplatelet drugs. Antiplatelet drugs that may be administered with the compositions of the invention include, but are not limited to, aspirin, dipyridamole (e.g., PERSANTINE™), and ticlopidine (e.g., TICLID™).

In specific embodiments, the use of anti-coagulants, thrombolytic and/or antiplatelet drugs in combination with Therapeutics of the invention is contemplated for the prevention, diagnosis, and/or treatment of thrombosis, arterial thrombosis, venous thrombosis, thromboembolism, pulmonary embolism, atherosclerosis, myocardial infarction, transient ischemic attack, unstable angina. In specific embodiments, the use of anticoagulants, thrombolytic drugs and/or antiplatelet drugs in combination with Therapeutics of the invention is contemplated for the prevention of occlusion of saphenous grafts, for reducing the risk of periprocedural thrombosis as might accompany angioplasty procedures, for reducing the risk of stroke in patients with atrial fibrillation including nonrheumatic atrial fibrillation, for reducing the risk of embolism associated with mechanical heart valves and or mitral valves disease. Other uses for the therapeutics of the invention, alone or in combination with antiplatelet, anticoagulant, and/or thrombolytic drugs, include, but are not limited to, the prevention of occlusions in extracorporeal devices (e.g., intravascular canulas, vascular access shunts in hemodialysis patients, hemodialysis machines, and cardiopulmonary bypass machines).

In certain embodiments, Therapeutics of the invention are administered in combination with antiretroviral agents, nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), and/or protease inhibitors (PIs). NRTIs that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, RETROVIR™ (zidovudine/AZT), VIDEX™ (didanosine/ddI), HIVID™. (zalcitabine/ddC), ZERIT™ (stavudine/d4T), EPIVIR™ (lamivudine/3TC), and COMBIVIR™ (zidovudine/lamivudine). NNRTIs that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, VIRAMUNE™ (nevirapine), RESCRIPTOR™ (delavirdine), and SUSTIVA™ (efavirenz). Protease inhibitors that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, CRIXIVAN™ (indinavir), NORVIR™ (ritonavir), INVIRASE™ (saquinavir), and VIRACEPT™ (nelfinavir). In a specific embodiment, antiretroviral agents, nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, and/or protease inhibitors may be used in any combination with Therapeutics of the invention to treat AIDS and/or to prevent or treat HIV infection.

Additional NRTIs include LODENOSINE™ (F-ddA; an acid-stable adenosine NRTI; Triangle/Abbott; COVIRACIL™ (emtricitabine/FTC; structurally related to lamivudine (3TC) but with 3- to 10-fold greater activity in vitro; Triangle/Abbott); dOTC (BCH-10652, also structurally related to lamivudine but retains activity against a substantial proportion of lamivudine-resistant isolates; Biochem Pharma); Adefovir (refused approval for anti-HIV therapy by FDA; Gilead Sciences); PREVEON® (Adefovir Dipivoxil, the active prodrug of adefovir; its active form is PMEA-pp); TENOFOVIR™ (bis-POC PMPA, a PMPA prodrug; Gilead); DAPD/DXG (active metabolite of DAPD; Triangle/Abbott); D-D4FC (related to 3TC, with activity against AZT/3TC-resistant virus); GW420867X (Glaxo Wellcome); ZIAGEN™ (abacavir/159U89; Glaxo Wellcome Inc.); CS-87 (3′azido-2′,3′-dideoxyuridine; WO 99/66936); and S-acyl-2-thioethyl (SATE)-bearing prodrug forms of β-L-FD4C and β-L-FddC (WO 98/17281).

Additional NNRTIs include COACTINON™ (Emivirine/MKC-442, potent NNRTI of the HEPT class; Triangle/Abbott); CAPRAVIRINE™ (AG-1549/S-1153, a next generation NNRTI with activity against viruses containing the K103N mutation; Agouron); PNU-142721 (has 20- to 50-fold greater activity than its predecessor delavirdine and is active against K103N mutants; Pharmacia & Upjohn); DPC-961 and DPC-963 (second-generation derivatives of efavirenz, designed to be active against viruses with the K103N mutation; DuPont); GW-420867X (has 25-fold greater activity than HBY097 and is active against K103N mutants; Glaxo Wellcome); CALANOLIDE A (naturally occurring agent from the latex tree; active against viruses containing either or both the Y181C and K103N mutations); and Propolis (WO 99/49830).

Additional protease inhibitors include LOPINAVIR™ (ABT378/r; Abbott Laboratories); BMS-232632 (an azapeptide; Bristol-Myres Squibb); TIPRANAVIR™ (PNU-140690, a non-peptic dihydropyrone; Pharmacia & Upjohn); PD-178390 (a nonpeptidic dihydropyrone; Parke-Davis); BMS 232632 (an azapeptide; Bristol-Myers Squibb); L-756,423 (an indinavir analog; Merck); DMP-450 (a cyclic urea compound; Avid & DuPont); AG-1776 (a peptidomimetic with in vitro activity against protease inhibitor-resistant viruses; Agouron); VX-175/GW-433908 (phosphate prodrug of amprenavir; Vertex & Glaxo Welcome); CGP61755 (Ciba); and AGENERASE™ (amprenavir; Glaxo Wellcome Inc.).

Additional antiretroviral agents include fusion inhibitors/gp41 binders. Fusion inhibitors/gp41 binders include T-20 (a peptide from residues 643-678 of the HIV gp41 transmembrane protein ectodomain which binds to gp41 in its resting state and prevents transformation to the fusogenic state; Trimeris) and T-1249 (a second-generation fusion inhibitor; Trimeris).

Additional antiretroviral agents include fusion inhibitors/chemokine receptor antagonists. Fusion inhibitors/chemokine receptor antagonists include CXCR4 antagonists such as AMD 3100 (a bicyclam), SDF-1 and its analogs, and ALX40-4C (a cationic peptide), T22 (an 18 amino acid peptide; Trimeris) and the T22 analogs T134 and T140; CCR5 antagonists such as RANTES (9-68), AOP-RANTES, NNY-RANTES, and TAK-779; and CCR5/CXCR4 antagonists such as NSC 651016 (a distamycin analog). Also included are CCR2B, CCR3, and CCR6 antagonists. Chemokine receptor agonists such as RANTES, SDF-1, MIP-1α, MIP-1β, etc., may also inhibit fusion.

Additional antiretroviral agents include integrase inhibitors. Integrase inhibitors include dicaffeoylquinic (DFQA) acids; L-chicoric acid (a dicaffeoyltartaric (DCTA) acid); quinalizarin (QLC) and related anthraquinones; ZINTEVIR™ (AR 177, an oligonucleotide that probably acts at cell surface rather than being a true integrase inhibitor; Arondex); and naphthols such as those disclosed in WO 98/50347.

Additional antiretroviral agents include hydroxyurea-like compounds such as BCX-34 (a purine nucleoside phosphorylase inhibitor; Biocryst); ribonucleotide reductase inhibitors such as DIDOX™ (Molecules for Health); inosine monophosphate dehydrogenase (IMPDH) inhibitors sucha as VX-497 (Vertex); and mycopholic acids such as CellCept (mycophenolate mofetil; Roche).

Additional antiretroviral agents include inhibitors of viral integrase, inhibitors of viral genome nuclear translocation such as arylene bis(methylketone) compounds; inhibitors of HIV entry such as AOP-RANTES, NNY-RANTES, RANTES-IgG fusion protein, soluble complexes of RANTES and glycosaminoglycans (GAG), and AMD-3100; nucleocapsid zinc finger inhibitors such as dithiane compounds; targets of HIV Tat and Rev; and pharmacoenhancers such as ABT-378.

Other antiretroviral therapies and adjunct therapies include cytokines and lymphokines such as MIP-1α, MIP-1β, SDF-1α, IL-2, PROLEUKIN™ (aldesleukin/L2-7001; Chiron), IL-4, IL-10, IL-12, and IL-13; interferons such as IFN-α2a; antagonists of TNFs, NFκB, GM-CSF, M-CSF, and IL-10; agents that modulate immune activation such as cyclosporin and prednisone; vaccines such as Remune™ (HIV Immunogen), APL 400-003 (Apollon), recombinant gp120 and fragments, bivalent (B/E) recombinant envelope glycoprotein, rgp120CM235, MN rgp120, SF-2 rgp120, gp120/soluble CD4 complex, Delta JR-FL protein, branched synthetic peptide derived from discontinuous gp120 C3/C4 domain, fusion-competent immunogens, and Gag, Pol, Nef, and Tat vaccines; gene-based therapies such as genetic suppressor elements (GSEs; WO 98/54366), and intrakines (genetically modified CC chemokines targetted to the ER to block surface expression of newly synthesized CCR5 (Yang et al., PNAS 94:11567-72 (1997); Chen et al., Nat. Med. 3:1110-16 (1997)); antibodies such as the anti-CXCR4 antibody 12G5, the anti-CCR5 antibodies 2D7, 5C7, PA8, PA9, PA10, PA11, PA12, and PA14, the anti-CD4 antibodies Q4120 and RPA-T4, the anti-CCR3 antibody 7B11, the anti-gp120 antibodies 17b, 48d, 447-52D, 257-D, 268-D and 50.1, anti-Tat antibodies, anti-TNF-α antibodies, and monoclonal antibody 33A; aryl hydrocarbon (AH) receptor agonists and antagonists such as TCDD, 3,3′,4,4′,5-pentachlorobiphenyl, 3,3′,4,4′-tetrachlorobiphenyl, and α-naphthoflavone (WO 98/30213); and antioxidants such as γ-L-glutamyl-L-cysteine ethyl ester (γ-GCE; WO 99/56764).

In a further embodiment, the Therapeutics of the invention are administered in combination with an antiviral agent. Antiviral agents that may be administered with the Therapeutics of the invention include, but are not limited to, acyclovir, ribavirin, amantadine, and remantidine.

In other embodiments, Therapeutics of the invention may be administered in combination with anti-opportunistic infection agents. Anti-opportunistic agents that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, TRIMETHOPRIM-SULFAMETHOXAZOLE™, DAPSONE™, PENTAMIDINE™, ATOVAQUONE™, ISONIAZID™, RIFAMPIN™, PYRAZINAMIDE™, ETHAMBUTOL™, RIFABUTIN™, CLARITHROMYCIN™, AZITHROMYCIN™, GANCICLOVIR™, FOSCARNET™, CIDOFOVIR™, FLUCONAZOLE™, ITRACONAZOLE™, KETOCONAZOLE™, ACYCLOVIR™, FAMCICOLVIR™, PYRIMETHAMINE™, LEUCOVORIN™, NEUPOGEN™ (filgrastim/G-CSF), and LEUKINE™ (sargramostim/GM-CSF). In a specific embodiment, Therapeutics of the invention are used in any combination with TRIMETHOPRIM-SULFAMETHOXAZOLE™, DAPSONE™, PENTAMIDINE™, and/or ATOVAQUONE™ to prophylactically treat or prevent an opportunistic Pneumocystis carinii pneumonia infection. In another specific embodiment, Therapeutics of the invention are used in any combination with ISONIAZID™, RIFAMPIN™, PYRAZINAMIDE™, and/or ETHAMBUTOL™ to prophylactically treat or prevent an opportunistic Mycobacterium avium complex infection. In another specific embodiment, Therapeutics of the invention are used in any combination with RIFABUTIN™, CLARITHROMYCIN™, and/or AZITHROMYCIN™ to prophylactically treat or prevent an opportunistic Mycobacterium tuberculosis infection. In another specific embodiment, Therapeutics of the invention are used in any combination with GANCICLOVIR™, FOSCARNET™, and/or CIDOFOVIR™ to prophylactically treat or prevent an opportunistic cytomegalovirus infection. In another specific embodiment, Therapeutics of the invention are used in any combination with FLUCONAZOLE™, ITRACONAZOLE™, and/or KETOCONAZOLE™ to prophylactically treat or prevent an opportunistic fungal infection. In another specific embodiment, Therapeutics of the invention are used in any combination with ACYCLOVIR™ and/or FAMCICOLVIR™ to prophylactically treat or prevent an opportunistic herpes simplex virus type I and/or type II infection. In another specific embodiment, Therapeutics of the invention are used in any combination with PYREMETHAMINE™ and/or LEUCOVORIN™ to prophylactically treat or prevent an opportunistic Toxoplasma gondii infection. In another specific embodiment, Therapeutics of the invention are used in any combination with LEUCOVORIN™ and/or NEUPOGEN™ to prophylactically treat or prevent an opportunistic bacterial infection.

In a further embodiment, the Therapeutics of the invention are administered in combination with an antibiotic agent. Antibiotic agents that may be administered with the Therapeutics of the invention include, but are not limited to, amoxicillin, beta-lactamases, aminoglycosides, beta-lactam (glycopeptide), beta-lactamases, Clindamycin, chloramphenicol, cephalosporins, ciprofloxacin, erythromycin, fluoroquinolones, macrolides, metronidazole, penicillins, quinolones, rapamycin, rifampin, streptomycin, sulfonamide, tetracyclines, trimethoprim, trimethoprim-sulfamethoxazole, and vancomycin.

In other embodiments, the Therapeutics of the invention are administered in combination with immunestimulants. Immunostimulants that may be administered in combination with the Therapeutics of the invention include, but are not limited to, levamisole (e.g., ERGAMISOL™), isoprinosine (e.g. INOSIPLEX™), interferons (e.g. interferon alpha), and interleukins (e.g., IL-2).

In other embodiments, Therapeutics of the invention are administered in combination with immunosuppressive agents. Immunosuppressive agents that may be administered in combination with the Therapeutics of the invention include, but are not limited to, steroids, cyclosporine, cyclosporine analogs, cyclophosphamide methylprednisone, prednisone, azathioprine, FK-506, 15-deoxyspergualin, and other immunosuppressive agents that act by suppressing the function of responding T cells. Other immunosuppressive agents that may be administered in combination with the Therapeutics of the invention include, but are not limited to, prednisolone, methotrexate, thalidomide, methoxsalen, rapamycin, leflunomide, mizoribine (BREDININ™), brequinar, deoxyspergualin, and azaspirane (SKF 105685), ORTHOCLONE OKT® 3 (muromonab-CD3), SANDIMMUNE™, NEORAL™, SANGDYA™ (cyclosporine), PROGRAF.® (FK506, tacrolimus), CELLCEPT® (mycophenolate motefil, of which the active metabolite is mycophenolic acid), IMURAN™ (azathioprine), glucocorticosteroids, adrenocortical steroids such as DELTASONE™ (prednisone) and HYDELTRASOL™ (prednisolone), FOLEX™ and MEXATE™ (methotrxate), OXSORALEN-ULTRA™ (methoxsalen) and RAPAMUNE™ (sirolimus). In a specific embodiment, immunosuppressants may be used to prevent rejection of organ or bone marrow transplantation.

In an additional embodiment, Therapeutics of the invention are administered alone or in combination with one or more intravenous immune globulin preparations. Intravenous immune globulin preparations that may be administered with the Therapeutics of the invention include, but not limited to, GAMMAR™, IVEEGAM™, SANDOGLOBULIN™, GAMMAGARD S/D™, ATGAM™ (antithymocyte glubulin), and GAMIMUNE™. In a specific embodiment, Therapeutics of the invention are administered in combination with intravenous immune globulin preparations in transplantation therapy (e.g., bone marrow transplant).

In certain embodiments, the Therapeutics of the invention are administered alone or in combination with an anti-inflammatory agent. Anti-inflammatory agents that may be administered with the Therapeutics of the invention include, but are not limited to, corticosteroids (e.g. betamethasone, budesonide, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone), nonsteroidal anti-inflammatory drugs (e.g., diclofenac, diflunisal, etodolac, fenoprofen, floctafenine, flurbiprofen, ibuprofen, indomethacin, ketoprofen, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, phenylbutazone, piroxicam, sulindac, tenoxicam, tiaprofenic acid, and tolmetin.), as well as antihistamines, aminoarylcarboxylic acid derivatives, arylacetic acid derivatives, arylbutyric acid derivatives, arylcarboxylic acids, arylpropionic acid derivatives, pyrazoles, pyrazolones, salicylic acid derivatives, thiazinecarboxamides, e-acetamidocaproic acid, S-adenosylmethionine, 3-amino-4-hydroxybutyric acid, amixetrine, bendazac, benzydamine, bucolome, difenpiramide, ditazol, emorfazone, guaiazulene, nabumetone, nimesulide, orgotein, oxaceprol, paranyline, perisoxal, pifoxime, proquazone, proxazole, and tenidap.

In an additional embodiment, the compositions of the invention are administered alone or in combination with an anti-angiogenic agent. Anti-angiogenic agents that may be administered with the compositions of the invention include, but are not limited to, Angiostatin (Entremed, Rockville, Md.), Troponin-1 (Boston Life Sciences, Boston, Mass.), anti-Invasive Factor, retinoic acid and derivatives thereof, paclitaxel (Taxol), Suramin, Tissue Inhibitor of Metalloproteinase-1, Tissue Inhibitor of Metalloproteinase-2, VEGI, Plasminogen Activator Inhibitor-1, Plasminogen Activator Inhibitor-2, and various forms of the lighter “d group” transition metals.

Lighter “d group” transition metals include, for example, vanadium, molybdenum, tungsten, titanium, niobium, and tantalum species. Such transition metal species may form transition metal complexes. Suitable complexes of the above-mentioned transition metal species include oxo transition metal complexes.

Representative examples of vanadium complexes include oxo vanadium complexes such as vanadate and vanadyl complexes. Suitable vanadate complexes include metavanadate and orthovanadate complexes such as, for example, ammonium metavanadate, sodium metavanadate, and sodium orthovanadate. Suitable vanadyl complexes include, for example, vanadyl acetylacetonate and vanadyl sulfate including vanadyl sulfate hydrates such as vanadyl sulfate mono- and trihydrates.

Representative examples of tungsten and molybdenum complexes also include oxo complexes. Suitable oxo tungsten complexes include tungstate and tungsten oxide complexes. Suitable tungstate complexes include ammonium tungstate, calcium tungstate, sodium tungstate dihydrate, and tungstic acid. Suitable tungsten oxides include tungsten (IV) oxide and tungsten (VI) oxide. Suitable oxo molybdenum complexes include molybdate, molybdenum oxide, and molybdenyl complexes. Suitable molybdate complexes include ammonium molybdate and its hydrates, sodium molybdate and its hydrates, and potassium molybdate and its hydrates. Suitable molybdenum oxides include molybdenum (VI) oxide, molybdenum (VI) oxide, and molybdic acid. Suitable molybdenyl complexes include, for example, molybdenyl acetylacetonate. Other suitable tungsten and molybdenum complexes include hydroxo derivatives derived from, for example, glycerol, tartaric acid, and sugars.

A wide variety of other anti-angiogenic factors may also be utilized within the context of the present invention. Representative examples include, but are not limited to, platelet factor 4; protamine sulphate; sulphated chitin derivatives (prepared from queen crab shells), (Murata et al., Cancer Res. 51:22-26, (1991)); Sulphated Polysaccharide Peptidoglycan Complex (SP-PG) (the function of this compound may be enhanced by the presence of steroids such as estrogen, and tamoxifen citrate); Staurosporine; modulators of matrix metabolism, including for example, proline analogs, cishydroxyproline, d,L-3,4-dehydroproline, Thiaproline, alpha,alpha-dipyridyl, aminopropionitrile fumarate; 4-propyl-5-(4-pyridinyl)-2(3H)-oxazolone; Methotrexate; Mitoxantrone; Heparin; Interferons; 2 Macroglobulin-serum; ChIMP-3 (Pavloff et al., J. Bio. Chem. 267:17321-17326, (1992)); Chymostatin (Tomkinson et al., Biochem J. 286:475-480, (1992)); Cyclodextrin Tetradecasulfate; Eponemycin; Camptothecin; Fumagillin (Ingber et al., Nature 348:555-557, (1990)); Gold Sodium Thiomalate (“GST”; Matsubara and Ziff, J. Clin. Invest. 79:1440-1446, (1987)); anticollagenase-serum; alpha2-antiplasmin (Holmes et al., J. Biol. Chem. 262(4):1659-1664, (1987)); Bisantrene (National Cancer Institute); Lobenzarit disodium (N-(2)-carboxyphenyl-4-chloroanthronilic acid disodium or “CCA”; (Takeuchi et al., Agents Actions 36:312-316, (1992)); and metalloproteinase inhibitors such as BB94.

Additional anti-angiogenic factors that may also be utilized within the context of the present invention include Thalidomide, (Celgene, Warren, N.J.); Angiostatic steroid; AGM-1470 (H. Brem and J. Folkman J Pediatr. Surg. 28:445-51 (1993)); an integrin alpha v beta 3 antagonist (C. Storgard et al., J. Clin. Invest. 103:47-54 (1999)); carboxynaminolmidazole; Carboxyamidotriazole (CAI) (National Cancer Institute, Bethesda, Md.); Conbretastatin A-4 (CA4P) (OXiGENE, Boston, Mass.); Squalamine (Magainin Pharmaceuticals, Plymouth Meeting, Pa.); TNP-470, (Tap Pharmaceuticals, Deerfield, Ill.); ZD-0101 AstraZeneca (London, UK); APRA (CT2584); Benefin, Byrostatin-1 (SC339555); CGP-41251 (PKC 412); CM101; Dexrazoxane (ICRF187); DMXAA; Endostatin; Flavopridiol; Genestein; GTE; ImmTher; Iressa (ZD1839); Octreotide (Somatostatin); Panretin; Penacillamine; Photopoint; PI-88; Prinomastat (AG-3340) Purlytin; Suradista (FCE26644); Tamoxifen (Nolvadex); Tazarotene; Tetrathiomolybdate; Xeloda (Capecitabine); and 5-Fluorouracil.

Anti-angiogenic agents that may be administered in combination with the compounds of the invention may work through a variety of mechanisms including, but not limited to, inhibiting proteolysis of the extracellular matrix, blocking the function of endothelial cell-extracellular matrix adhesion molecules, by antagonizing the function of angiogenesis inducers such as growth factors, and inhibiting integrin receptors expressed on proliferating endothelial cells. Examples of anti-angiogenic inhibitors that interfere with extracellular matrix proteolysis and which may be administered in combination with the compositions of the invention include, but are not limited to, AG-3340 (Agouron, La Jolla, Calif.), BAY-12-9566 (Bayer, West Haven, Conn.), BMS-275291 (Bristol Myers Squibb, Princeton, N.J.), CGS-27032A (Novartis, East Hanover, N.J.), Marimastat (British Biotech, Oxford, UK), and Metastat (Aeterna, St-Foy, Quebec). Examples of anti-angiogenic inhibitors that act by blocking the function of endothelial cell-extracellular matrix adhesion molecules and which may be administered in combination with the compositions of the invention include, but are not limited to, EMD-121974 (Merck KcgaA Darmstadt, Germany) and Vitaxin (Ixsys, La Jolla, Calif./Medimmune, Gaithersburg, Md.). Examples of anti-angiogenic agents that act by directly antagonizing or inhibiting angiogenesis inducers and which may be administered in combination with the compositions of the invention include, but are not limited to, Angiozyme (Ribozyme, Boulder, Colo.), Anti-VEGF antibody (Genentech, S. San Francisco, Calif.), PTK-787/ZK-225846 (Novartis, Basel, Switzerland), SU-101 (Sugen, S. San Francisco, Calif.), SU-5416 (Sugen/Pharmacia Upjohn, Bridgewater, N.J.), and SU-6668 (Sugen). Other anti-angiogenic agents act to indirectly inhibit angiogenesis. Examples of indirect inhibitors of angiogenesis which may be administered in combination with the compositions of the invention include, but are not limited to, IM-862 (Cytran, Kirkland, Wash.), Interferon-alpha, IL-12 (Roche, Nutley, N.J.), and Pentosan polysulfate (Georgetown University, Washington, D.C.).

In particular embodiments, the use of compositions of the invention in combination with anti-angiogenic agents is contemplated for the treatment, prevention, and/or amelioration of an autoimmune disease, such as for example, an autoimmune disease described herein.

In a particular embodiment, the use of compositions of the invention in combination with anti-angiogenic agents is contemplated for the treatment, prevention, and/or amelioration of arthritis. In a more particular embodiment, the use of compositions of the invention in combination with anti-angiogenic agents is contemplated for the treatment, prevention, and/or amelioration of rheumatoid arthritis.

In another embodiment, the polynucleotides encoding a polypeptide of the present invention are administered in combination with an angiogenic protein, or polynucleotides encoding an angiogenic protein. Examples of angiogenic proteins that may be administered with the compositions of the invention include, but are not limited to, acidic and basic fibroblast growth factors, VEGF-1, VEGF-2, VEGF-3, epidermal growth factor alpha and beta, platelet-derived endothelial cell growth factor, platelet-derived growth factor, tumor necrosis factor alpha, hepatocyte growth factor, insulin-like growth factor, colony stimulating factor, macrophage colony stimulating factor, granulocyte/macrophage colony stimulating factor, and nitric oxide synthase.

In additional embodiments, compositions of the invention are administered in combination with a chemotherapeutic agent. Chemotherapeutic agents that may be administered with the Therapeutics of the invention include, but are not limited to alkylating agents such as nitrogen mustards (for example, Mechlorethamine, cyclophosphamide, Cyclophosphamide Ifosfamide, Melphalan (L-sarcolysin), and Chlorambucil), ethylenimines and methylmelamines (for example, Hexamethylmelamine and Thiotepa), alkyl sulfonates (for example, Busulfan), nitrosoureas (for example, Cammustine (BCNU), Lomustine (CCNU), Semustine (methyl-CCNU), and Streptozocin (streptozotocin)), triazenes (for example, Dacarbazine (DTIC; dimethyltriazenoimidazolecarboxamide)), folic acid analogs (for example, Methotrexate (amethopterin)), pyrimidine analogs (for example, Fluorouacil (5-fluorouracil; 5-FU), Floxuridine (fluorodeoxyuridine; FudR), and Cytarabine (cytosine arabinoside)), purine analogs and related inhibitors (for example, Mercaptopurine (6-mercaptopurine; 6-MP), Thioguanine (6-thioguanine; TG), and Pentostatin (2′-deoxycoformycin)), vinca alkaloids (for example, Vinblastine (VLB, vinblastine sulfate)) and Vincristine (vincristine sulfate)), epipodophyllotoxins (for example, Etoposide and Teniposide), antibiotics (for example, Dactinomycin (actinomycin D), Daunorubicin (daunomycin; rubidomycin), Doxorubicin, Bleomycin, Plicamycin (mithramycin), and Mitomycin (mitomycin C), enzymes (for example, L-Asparaginase), biological response modifiers (for example, Interferon-alpha and interferon-alpha-2b), platinum coordination compounds (for example, Cisplatin (cis-DDP) and Carboplatin), anthracenedione (Mitoxantrone), substituted ureas (for example, Hydroxyurea), methylhydrazine derivatives (for example, Procarbazine (N-methylhydrazine; M1H), adrenocorticosteroids (for example, Prednisone), progestins (for example, Hydroxyprogesterone caproate, Medroxyprogesterone, Medroxyprogesterone acetate, and Megestrol acetate), estrogens (for example, Diethylstilbestrol (DES), Diethylstilbestrol diphosphate, Estradiol, and Ethinyl estradiol), antiestrogens (for example, Tamoxifen), androgens (Testosterone proprionate, and Fluoxymesterone), antiandrogens (for example, Flutamide), gonadotropin-releasing hormone analogs (for example, Leuprolide), other hormones and hormone analogs (for example, methyltestosterone, estramustine, estramustine phosphate sodium, chlorotrianisene, and testolactone), and others (for example, dicarbazine, glutamic acid, and mitotane).

In one embodiment, the compositions of the invention are administered in combination with one or more of the following drugs: infliximab (also known as Remicade™ Centocor, Inc.), Trocade (Roche, RO-32-3555), Leflunomide (also known as Arava™ from Hoechst Marion Roussel), Kineret™ (an IL-1 Receptor antagonist also known as Anakinra from Amgen, Inc.)

In a specific embodiment, compositions of the invention are administered in combination with CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) or combination of one or more of the components of CHOP. In one embodiment, the compositions of the invention are administered in combination with anti-CD20 antibodies, human monoclonal anti-CD20 antibodies. In another embodiment, the compositions of the invention are administered in combination with anti-CD20 antibodies and CHOP, or anti-CD20 antibodies and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. In a specific embodiment, compositions of the invention are administered in combination with Rituximab. In a further embodiment, compositions of the invention are administered with Rituximab and CHOP, or Rituximab and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. In a specific embodiment, compositions of the invention are administered in combination with tositumomab. In a further embodiment, compositions of the invention are administered with tositumomab and CHOP, or tositumomab and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. The anti-CD20 antibodies may optionally be associated with radioisotopes, toxins or cytotoxic prodrugs.

In another specific embodiment, the compositions of the invention are administered in combination Zevalin™. In a further embodiment, compositions of the invention are administered with Zevalin™ and CHOP, or Zevalin™ and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. Zevalin™ may be associated with one or more radisotopes. Particularly preferred isotopes are ⁹⁰Y and ¹¹¹In.

In an additional embodiment, the Therapeutics of the invention are administered in combination with cytokines. Cytokines that may be administered with the Therapeutics of the invention include, but are not limited to, IL2, IL3, IL4, IL5, IL6, IL7, IL10, IL12, IL13, IL15, anti-CD40, CD40L, IFN-gamma and TNF-alpha. In another embodiment, Therapeutics of the invention may be administered with any interleukin, including, but not limited to, IL-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, and IL-21.

In one embodiment, the Therapeutics of the invention are administered in combination with members of the TNF family. TNF, TNF-related or TNF-like molecules that may be administered with the Therapeutics of the invention include, but are not limited to, soluble forms of TNF-alpha, lymphotoxin-alpha (LT-alpha, also known as TNF-beta), LT-beta (found in complex heterotrimer LT-alpha2-beta), OPGL, FasL, CD27L, CD30L, CD40L, 4-IBBL, DcR3, OX40L, TNF-gamma (International Publication No. WO 96/14328), AIM-I (International Publication No. WO 97/33899), endokine-alpha (International Publication No. WO 98/07880), OPG, and neutrokine-alpha (International Publication No. WO 98/18921, OX40, and nerve growth factor (NGF), and soluble forms of Fas, CD30, CD27, CD40 and 4-IBB, TR2 (International Publication No. WO 96/34095), DR3 (International Publication No. WO 97/33904), DR4 (International Publication No. WO 98/32856), TR5 (International Publication No. WO 98/30693), TRANK, TR9 (International Publication No. WO 98/56892), TR10 (International Publication No. WO 98/54202), 312C2 (International Publication No. WO 98/06842), and TR12, and soluble forms CD154, CD70, and CD153.

In an additional embodiment, the Therapeutics of the invention are administered in combination with angiogenic proteins. Angiogenic proteins that may be administered with the Therapeutics of the invention include, but are not limited to, Glioma Derived Growth Factor (GDGF), as disclosed in European Patent Number EP-399816; Platelet Derived Growth Factor-A (PDGF-A), as disclosed in European Patent Number EP-682110; Platelet Derived Growth Factor-B (PDGF-B), as disclosed in European Patent Number EP-282317; Placental Growth Factor (PlGF), as disclosed in International Publication Number WO 92/06194; Placental Growth Factor-2 (PlGF-2), as disclosed in Hauser et al., Growth Factors, 4:259-268 (1993); Vascular Endothelial Growth Factor (VEGF), as disclosed in International Publication Number WO 90/13649; Vascular Endothelial Growth Factor-A (VEGF-A), as disclosed in European Patent Number EP-506477; Vascular Endothelial Growth Factor-2 (VEGF-2), as disclosed in International Publication Number WO 96/39515; Vascular Endothelial Growth Factor B (VEGF-3); Vascular Endothelial Growth Factor B-186 (VEGF-B186), as disclosed in International Publication Number WO 96/26736; Vascular Endothelial Growth Factor-D (VEGF-D), as disclosed in International Publication Number WO 98/02543; Vascular Endothelial Growth Factor-D (VEGF-D), as disclosed in International Publication Number WO 98/07832; and Vascular Endothelial Growth Factor-E (VEGF-E), as disclosed in German Patent Number DE19639601. The above mentioned references are herein incorporated by reference in their entireties.

In an additional embodiment, the Therapeutics of the invention are administered in combination with Fibroblast Growth Factors. Fibroblast Growth Factors that may be administered with the Therapeutics of the invention include, but are not limited to, FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7, FGF-8, FGF-9, FGF-10, FGF-11, FGF-12, FGF-13, FGF-14, and FGF-15.

In an additional embodiment, the Therapeutics of the invention are administered in combination with hematopoietic growth factors. Hematopoietic growth factors that may be administered with the Therapeutics of the invention include, but are not limited to, granulocyte macrophage colony stimulating factor (GM-CSF) (sargramostim, LEUKINE™, PROKINE™), granulocyte colony stimulating factor (G-CSF) (filgrastim, NEUPOGEN™), macrophage colony stimulating factor (M-CSF, CSF-1) erythropoietin (epoetin alfa, EPOGEN™, PROCRIT™), stem cell factor (SCF, c-kit ligand, steel factor), megakaryocyte colony stimulating factor, PIXY321 (a GMCSF/IL-3 fusion protein), interleukins, especially any one or more of IL-1 through IL-12, interferon-gamma, or thrombopoietin.

In certain embodiments, Therapeutics of the present invention are administered in combination with adrenergic blockers, such as, for example, acebutolol, atenolol, betaxolol, bisoprolol, carteolol, labetalol, metoprolol, nadolol, oxprenolol, penbutolol, pindolol, propranolol, sotalol, and timolol.

In another embodiment, the Therapeutics of the invention are administered in combination with an antiarrhythmic drug (e.g., adenosine, amidoarone, bretylium, digitalis, digoxin, digitoxin, diliazem, disopyramide, esmolol, flecainide, lidocaine, mexiletine, moricizine, phenyloin, procainamide, N-acetyl procainamide, propafenone, propranolol, quinidine, sotalol, tocainide, and verapamil).

In another embodiment, the Therapeutics of the invention are administered in combination with diuretic agents, such as carbonic anhydrase-inhibiting agents (e.g., acetazolamide, dichlorphenamide, and methazolamide), osmotic diuretics (e.g., glycerin, isosorbide, mannitol, and urea), diuretics that inhibit Na⁺—K⁺-2Cl⁻ symport (e.g., furosemide, bumetanide, azosemide, piretanide, tripamide, ethacrynic acid, muzolimine, and torsemide), thiazide and thiazide-like diuretics (e.g., bendroflumethiazide, benzthiazide, chlorothiazide, hydrochlorothiazide, hydroflumethiazide, methyclothiazide, polythiazide, trichormethiazide, chlorthalidone, indapamide, metolazone, and quinethazone), potassium sparing diuretics (e.g., amiloride and triamterene), and mineralcorticoid receptor antagonists (e.g., spironolactone, canrenone, and potassium canrenoate).

In one embodiment, the Therapeutics of the invention are administered in combination with treatments for endocrine and/or hormone imbalance disorders. Treatments for endocrine and/or hormone imbalance disorders include, but are not limited to, ¹²⁷I, radioactive isotopes of iodine such as ¹³¹I and ¹²³I; recombinant growth hormone, such as HUMATROPE™ (recombinant somatropin); growth hormone analogs such as PROTROPIN™ (somatrem); dopamine agonists such as PARLODEL™ (bromocriptine); somatostatin analogs such as SANDOSTATIN™ (octreotide); gonadotropin preparations such as PREGNYL™, A.P.L.™ and PROFASI™ (chorionic gonadotropin (CG)), PERGONAL™ (menotropins), and METRODIN™ (urofollitropin (uFSH)); synthetic human gonadotropin releasing hormone preparations such as FACTREL™ and LUTREPULSE™ (gonadorelin hydrochloride); synthetic gonadotropin agonists such as LUPRON™ (leuprolide acetate), SUPPRELIN™ (histrelin acetate), SYNAREL™ (nafarelin acetate), and ZOLADEX™ (goserelin acetate); synthetic preparations of thyrotropin-releasing hormone such as RELEFACT TRH™ and THYPINONE™ (protirelin); recombinant human TSH such as THYROGEN™; synthetic preparations of the sodium salts of the natural isomers of thyroid hormones such as L-T₄™, SYNTHROID™ and LEVOTHROID™ (levothyroxine sodium), L-T₃™, CYTOMEL™ and TRIOSTAT™ (liothyroine sodium), and THYROLAR™ (liotrix); antithyroid compounds such as 6-n-propylthiouracil (propylthiouracil), 1-methyl-2-mercaptoimidazole and TAPAZOLE™ (methimazole), NEO-MERCAZOLE™ (carbimazole); beta-adrenergic receptor antagonists such as propranolol and esmolol; Ca²⁺ channel blockers; dexamethasone and iodinated radiological contrast agents such as TELEPAQUE™ (iopanoic acid) and ORAGRAFIN™ (sodium ipodate).

Additional treatments for endocrine and/or hormone imbalance disorders include, but are not limited to, estrogens or congugated estrogens such as ESTRACE™ (estradiol), ESTINYL™ (ethinyl estradiol), PREMARIN™, ESTRATAB™, ORTHO-EST™, OGEN™ and estropipate (estrone), ESTROVIS™ (quinestrol), ESTRADERM™ (estradiol), DELESTROGEN™ and VALERGEN™ (estradiol valerate), DEPO-ESTRADIOL CYPIONATE™ and ESTROJECT LA™ (estradiol cypionate); antiestrogens such as NOLVADEX™ (tamoxifen), SEROPHENE™ and CLOMID™ (clomiphene); progestins such as DURALUTIN™ (hydroxyprogesterone caproate), MPA™ and DEPO-PROVERA™ (medroxyprogesterone acetate), PROVERA™ and CYCRIN™ (MPA), MEGACE™ (megestrol acetate), NORLUTIN™ (norethindrone), and NORLUTATE™ and AYGESTIN™ (norethindrone acetate); progesterone implants such as NORPLANT SYSTEM™ (subdermal implants of norgestrel); antiprogestins such as RU 486™ (mifcpristone); hormonal contraceptives such as ENOVID™ (norethynodrel plus mestranol), PROGESTASERT™ (intrauterine device that releases progesterone), LOESTRIN™, BREVICON™, MODICON™, GENORA™, NELONA™, NORINYL™, OVACON-35™ and OVACON-50™ (ethinyl estradiol/norethindrone), LEVLEN™, NORDETTE™, TR1-LEVLEN™ and TRIPHASIL-21™ (ethinyl estradiol/levonorgestrel) LO/OVRAL™ and OVRAL™ (ethinyl estradiol/norgestrel), DEMULEN™ (ethinyl estradiol/ethynodiol diacetate), NORINYL™, ORTHO-NOVUM™, NORETHIN™, GENORA™, and NELOVA™ (norethindrone/mestranol), DESOGEN™ and ORTHO-CEPT™ (ethinyl estradiol/desogestrel), ORTHO-CYCLEN™ and ORTHO-TRICYCLEN™ (ethinyl estradiol/norgestimate), MICRONOR™ and NOR-QD™ (norethindrone), and OVRETTE™ (norgestrel).

Additional treatments for endocrine and/or hormone imbalance disorders include, but are not limited to, testosterone esters such as methenolone acetate and testosterone undecanoate; parenteral and oral androgens such as TESTOJECT-50™ (testosterone), TESTEX™ (testosterone propionate), DELATESTRYL™ (testosterone enanthate), DEPO-TESTOSTERONE™ (testosterone cypionate), DANOCRINE™ (danazol), HALOTESTIN™ (fluoxymesterone), ORETON METHYL™, TESTRED™ and VIRILON™ (methyltestosterone), and OXANDRIN™ (oxandrolone); testosterone transdermal systems such as TESTODERM™; androgen receptor antagonist and 5-alpha-reductase inhibitors such as ANDROCUR™ (cyproterone acetate), EULEXIN™ (flutamide), and PROSCAR™ (finasteride); adrenocorticotropic hormone preparations such as CORTROSYN™ (cosyntropin); adrenocortical steroids and their synthetic analogs such as ACLOVATE™ (alclometasone dipropionate), CYCLOCORT™ (amcinonide), BECLOVENT™ and VANCERIL™ (beclomethasone dipropionate), CELESTONE™ (betamethasone), BENISONE™ and UTICORT™ (betamethasone benzoate), DIPROSONE™ (betamethasone dipropionate), CELESTONE PHOSPHATE™ (betamethasone sodium phosphate), CELESTONE SOLUSPAN™ (betamethasone sodium phosphate and acetate), BETA-VAL™ and VALISONE™ (betamethasone valerate), TEMOVATE™ (clobetasol propionate), CLODERM™ (clocortolone pivalate), CORTEF™ and HYDROCORTONE™ (cortisol (hydrocortisone)), HYDROCORTONE ACETATE™ (cortisol (hydrocortisone) acetate), LOCOID™ (cortisol (hydrocortisone) butyrate), HYDROCORTONE PHOSPHATE™ (cortisol (hydrocortisone) sodium phosphate), A-HYDROCORT™ and SOLU CORTEF™ (cortisol (hydrocortisone) sodium succinate), WESTCORT™ (cortisol (hydrocortisone) valerate), CORTISONE ACETATE™ (cortisone acetate), DESOWEN™ and TRIDESILON™ (desonide), TOPICORT™ (desoximetasone), DECADRON™ (dexamethasone), DECADRON LA™ (dexamethasone acetate), DECADRON PHOSPHATE™ and HEXADROL PHOSPHATE™ (dexametliasone sodium phosphate), FLORONE™ and MAXIFLOR™ (diflorasone diacetate), FLORINEF ACETATE™ (fludrocortisone acetate), AEROBID™ and NASALIDE™ (flunisolide), FLUONID™ and SYNALAR™ (fluocinolone acetonide), LIDEX™ (fluocinonide), FLUOR-OP™ and FML™ (fluorometholone), CORDRAN™ (flurandrenolide), HALOG™ (halcinonide), HMS LIZUIFILM™ (medrysone), MEDROL™ (methylprednisolone), DEPO-MEDROL™ and MEDROL ACETATE™ (methylprednisone acetate), A-METHAPRED™ and SOLUMEDROL™ (methylprednisolone sodium succinate), ELOCON™ (mometasone furoate), HALDRONE™ (paramethasone acetate), DELTA-CORTEF™ (prednisolone), ECONOPRED™ (prednisolone acetate), HYDELTRASOL™ (prednisolone sodium phosphate), HYDELTRA-T.B.A™ (prednisolone tebutate), DELTASONE™ (prednisone), ARISTOCORT™ and KENACORT™ (triamcinolone), KENALOG™ (triamcinolone acetonide), ARISTOCORT™ and KENACORT DIACETATE™ (triamcinolone diacetate), and ARISTOSPAN™ (triamcinolone hexacetonide); inhibitors of biosynthesis and action of adrenocortical steroids such as CYTADREN™ (aminoglutethimide), NIZORAL™ (ketoconazole), MODRASTANE™ (trilostane), and METOPIRONE™ (metyrapone); bovine, porcine or human insulin or mixtures thereof; insulin analogs; recombinant human insulin such as HUMULIN™ and NOVOLIN™; oral hypoglycemic agents such as ORAMIDE™ and ORINASE™ (tolbutamide), DIABINESE™ (chlorpropamide), TOLAMIDE™ and TOLINASE™ (tolazamide), DYMELOR™ (acetohexamide), glibenclamide, MICRONASE™, DIBETA™ and GLYNASE™ (glyburide), GLUCOTROL™ (glipizide), and DIAMICRON™ (gliciazide), GLUCOPHAGE™ (metformin), ciglitazone, pioglitazone, and alpha-glucosidase inhibitors; bovine or porcine glucagon; somatostatins such as SANDOSTATIN™ (octreotide); and diazoxides such as PROGLYCEM™ (diazoxide).

In one embodiment, the Therapeutics of the invention are administered in combination with treatments for uterine motility disorders. Treatments for uterine motility disorders include, but are not limited to, estrogen drugs such as conjugated estrogens (e.g., PREMARIN® and ESTRATAB®), estradiols (e.g., CLIMARA® and ALORA®), estropipate, and chlorotrianisene; progestin drugs (e.g., AMEN® (medroxyprogesterone), MICRONOR® (norethidrone acetate), PROMETRIUM® progesterone, and megestrol acetate); and estrogen/progesterone combination therapies such as, for example, conjugated estrogens/medroxyprogesterone (e.g., PREMPRO™ and PREMPHASE®) and norethindrone acetate/ethinyl estsradiol (e.g., FEMHRT™).

In an additional embodiment, the Therapeutics of the invention are administered in combination with drugs effective in treating iron deficiency and hypochromic anemias, including but not limited to, ferrous sulfate (iron sulfate, FEOSOL™), ferrous fumarate (e.g., FEOSTAT™), ferrous gluconate (e.g., FERGON™), polysaccharide-iron complex (e.g., NIFEREX™), iron dextran injection (e.g., INFED™), cupric sulfate, pyroxidine, riboflavin, Vitamin B₁₂, cyancobalamin injection (e.g., REDISOL™, RUBRAMIN PC™), hydroxocobalamin, folic acid (e.g., FOLVITE™), leucovorin (folinic acid, 5-CHOH4PteGlu, citrovorum factor) or WELLCOVORIN (Calcium salt of leucovorin), transferrin or ferritin.

In certain embodiments, the Therapeutics of the invention are administered in combination with agents used to treat psychiatric disorders. Psychiatric drugs that may be administered with the Therapeutics of the invention include, but are not limited to, antipsychotic agents (e.g., chlorpromazine, chlorprothixene, clozapine, fluphenazine, haloperidol, loxapine, mesoridazine, molindone, olanzapine, perphenazine, pimozide, quetiapine, risperidone, thioridazine, thiothixene, trifluoperazine, and triflupromazine), antimanic agents (e.g., carbamazepine, divalproex sodium, lithium carbonate, and lithium citrate), antidepressants (e.g., amitriptyline, amoxapine, bupropion, citalopram, clomipramine, desipramine, doxepin, fluvoxamine, fluoxetine, imipramine, isocarboxazid, maprotiline, mirtazapine, nefazodone, nortriptyline, paroxetine, phenelzine, protriptyline, sertraline, tranylcypromine, trazodone, trimipramine, and venlafaxine), antianxiety agents (e.g., alprazolam, buspirone, chlordiazepoxide, clorazepate, diazepam, halazepam, lorazepam, oxazepam, and prazepam), and stimulants (e.g., d-amphetamine, methylphenidate, and pemoline).

In other embodiments, the Therapeutics of the invention are administered in combination with agents used to treat neurological disorders. Neurological agents that may be administered with the Therapeutics of the invention include, but are not limited to, antiepileptic agents (e.g., carbamazepine, clonazepam, ethosuximide, phenobarbital, phenyloin, primidone, valproic acid, divalproex sodium, felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine, tiagabine, topiramate, zonisamide, diazepam, lorazepam, and clonazepam), antiparkinsonian agents (e.g., levodopa/carbidopa, selegiline, amantidine, bromocriptine, pergolide, ropinirole, pramipexole, benztropine; biperiden; ethopropazine; procyclidine; trihexyphenidyl, tolcapone), and ALS therapeutics (e.g. riluzole).

In another embodiment, Therapeutics of the invention are administered in combination with vasodilating agents and/or calcium channel blocking agents. Vasodilating agents that may be administered with the Therapeutics of the invention include, but are not limited to, Angiotensin Converting Enzyme (ACE) inhibitors (e.g., papaverine, isoxsuprine, benazepril, captopril, cilazapril, enalapril, enalaprilat, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, spirapril, trandolapril, and nyiidrin), and nitrates (e.g., isosorbide dinitrate, isosorbide mononitrate, and nitroglycerin). Examples of calcium channel blocking agents that may be administered in combination with the Therapeutics of the invention include, but are not limited to amlodipine, bepridil, diltiazem, felodipine, flunarizine, isradipine, nicardipine, nifedipine, nimodipine, and verapamil.

In certain embodiments, the Therapeutics of the invention are administered in combination with treatments for gastrointestinal disorders. Treatments for gastrointestinal disorders that may be administered with the Therapeutic of the invention include, but are not limited to, H₂ histamine receptor antagonists (e.g., TAGAMET™ (cimetidine), ZANTAC™ (ranitidine), PEPCID™ (famotidine), and AXID™ (nizatidine)); inhibitors of H⁺, K⁺ ATPase (e.g., PREVACID™ (lansoprazole) and PRILOSEC™ (omeprazole)); Bismuth compounds (e.g., PEPTO-BISMOL™ (bismuth subsalicylate) and DE-NOL™ (bismuth subcitrate)); various antacids; sucralfate; prostaglandin analogs (e.g. CYTOTEC™ (misoprostol)); muscarinic cholinergic antagonists; laxatives (e.g., surfactant laxatives, stimulant laxatives, saline and osmotic laxatives); antidiarrheal agents (e.g., LOMOTIL™ (diphenoxylate), MOTOFEN™ (diphenoxin), and IMODIUM™ (loperamide hydrochloride)), synthetic analogs of somatostatin such as SANDOSTATIN™ (octreotide), antiemetic agents (e.g., ZOFRAN™ (ondansetron), KYTRIL™ (granisetron hydrochloride), tropisetron, dolasetron, metoclopramide, chlorpromazine, perphenazine, prochlorperazine, promethazine, thiethylperazine, triflupromazine, domperidone, haloperidol, droperidol, trimethobenzamide, dexamethasone, methylprednisolone, dronabinol, and nabilone); D2 antagonists (e.g., metoclopramide, trimethobenzamide and chlorpromazine); bile salts; chenodeoxycholic acid; ursodeoxycholic acid; and pancreatic enzyme preparations such as pancreatin and pancrelipase.

In additional embodiments, the Therapeutics of the invention are administered in combination with other therapeutic or prophylactic regimens, such as, for example, radiation therapy.

Example 14 Method of Treating Decreased Levels of the Polypeptide

The present invention relates to a method for treating an individual in need of an increased level of a polypeptide of the invention in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of an agonist of the invention (including polypeptides of the invention). Moreover, it will be appreciated that conditions caused by a decrease in the standard or normal expression level of a polypeptide of the present invention in an individual can be treated by administering the agonist or antagonist of the present invention. Thus, the invention also provides a method of treatment of an individual in need of an increased level of the polypeptide comprising administering to such an individual a Therapeutic comprising an amount of the agonist or antagonist to increase the activity level of the polypeptide in such an individual.

For example, a patient with decreased levels of a polypeptide receives a daily dose 0.1-100 ug/kg of the agonist or antagonist for six consecutive days. The exact details of the dosing scheme, based on administration and formulation, are provided in Example 13.

Example 15 Method of Treating Increased Levels of the Polypeptide

The present invention also relates to a method of treating an individual in need of a decreased level of a polypeptide of the invention in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of an antagonist of the invention (including polypeptides and antibodies of the invention).

In one example, antisense technology is used to inhibit production of a polypeptide of the present invention. This technology is one example of a method of decreasing levels of a polypeptide, due to a variety of etiologies, such as cancer.

For example, a patient diagnosed with abnormally increased levels of a polypeptide is administered intravenously antisense polynucleotides at 0.5, 1.0, 1.5, 2.0 and 3.0 mg/kg day for 21 days. This treatment is repeated after a 7-day rest period if the treatment was well tolerated. The antisense polynucleotides of the present invention can be formulated using techniques and formulations described herein (e.g. see Example 13), or otherwise known in the art.

Example 16 Method of Treatment Using Gene Therapy-Ex Vivo

One method of gene therapy transplants fibroblasts, which are capable of expressing a polypeptide, onto a patient. Generally, fibroblasts are obtained from a subject by skin biopsy. The resulting tissue is placed in tissue-culture medium and separated into small pieces. Small chunks of the tissue are placed on a wet surface of a tissue culture flask, approximately ten pieces are placed in each flask. The flask is turned upside down, closed tight and left at room temperature over night. After 24 hours at room temperature, the flask is inverted and the chunks of tissue remain fixed to the bottom of the flask and fresh media (e.g., Ham's F12 media, with 10% FBS, penicillin and streptomycin) is added. The flasks are then incubated at 37 degree C. for approximately one week.

At this time, fresh media is added and subsequently changed every several days. After an additional two weeks in culture, a monolayer of fibroblasts emerge. The monolayer is trypsinized and scaled into larger flasks.

pMV-7 (Kirschmeier, P. T. et al., DNA, 7:219-25 (1988)), flanked by the long terminal repeats of the Moloney murine sarcoma virus, is digested with EcoRI and HindIII and subsequently treated with calf intestinal phosphatase. The linear vector is fractionated on agarose gel and purified, using glass beads.

The cDNA encoding a polypeptide of the present invention can be amplified using PCR primers which correspond to the 5′ and 3′ end sequences respectively as set forth in Example 1 using primers and having appropriate restriction sites and initiation/stop codons, if necessary. Preferably, the 5′ primer contains an EcoRI site and the 3′ primer includes a HindIII site. Equal quantities of the Moloney murine sarcoma virus linear backbone and the amplified EcoRI and HindIII fragment are added together, in the presence of T4 DNA ligase. The resulting mixture is maintained under conditions appropriate for ligation of the two fragments. The ligation mixture is then used to transform bacteria HB101, which are then plated onto agar containing kanamycin for the purpose of confirming that the vector has the gene of interest properly inserted.

The amphotropic pA317 or GP+am12 packaging cells are grown in tissue culture to confluent density in Dulbecco's Modified Eagles Medium (DMEM) with 10% calf serum (CS), penicillin and streptomycin. The MSV vector containing the gene is then added to the media and the packaging cells transduced with the vector. The packaging cells now produce infectious viral particles containing the gene (the packaging cells are now referred to as producer cells).

Fresh media is added to the transduced producer cells, and subsequently, the media is harvested from a 10 cm plate of confluent producer cells. The spent media, containing the infectious viral particles, is filtered through a millipore filter to remove detached producer cells and this media is then used to infect fibroblast cells. Media is removed from a sub-confluent plate of fibroblasts and quickly replaced with the media from the producer cells. This media is removed and replaced with fresh media. If the titer of virus is high, then virtually all fibroblasts will be infected and no selection is required. If the titer is very low, then it is necessary to use a retroviral vector that has a selectable marker, such as neo or his. Once the fibroblasts have been efficiently infected, the fibroblasts are analyzed to determine whether protein is produced.

The engineered fibroblasts are then transplanted onto the host, either alone or after having been grown to confluence on cytodex 3 microcarrier beads.

Example 17 Gene Therapy Using Endogenous Genes Corresponding to Polynucleotides of the Invention

Another method of gene therapy according to the present invention involves operably associating the endogenous polynucleotide sequence of the invention with a promoter via homologous recombination as described, for example, in U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication NO: WO 96/29411, published Sep. 26, 1996; International Publication NO: WO 94/12650, published Aug. 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA, 86:8932-8935 (1989); and Zijlstra et al., Nature, 342:435-438 (1989). This method involves the activation of a gene which is present in the target cells, but which is not expressed in the cells, or is expressed at a lower level than desired.

Polynucleotide constructs are made which contain a promoter and targeting sequences, which are homologous to the 5′ non-coding sequence of endogenous polynucleotide sequence, flanking the promoter. The targeting sequence will be sufficiently near the 5′ end of the polynucleotide sequence so the promoter will be operably linked to the endogenous sequence upon homologous recombination. The promoter and the targeting sequences can be amplified using PCR. Preferably, the amplified promoter contains distinct restriction enzyme sites on the 5′ and 3′ ends. Preferably, the 3′ end of the first targeting sequence contains the same restriction enzyme site as the 5′ end of the amplified promoter and the 5′ end of the second targeting sequence contains the same restriction site as the 3′ end of the amplified promoter.

The amplified promoter and the amplified targeting sequences are digested with the appropriate restriction enzymes and subsequently treated with calf intestinal phosphatase. The digested promoter and digested targeting sequences are added together in the presence of T4 DNA ligase. The resulting mixture is maintained under conditions appropriate for ligation of the two fragments. The construct is size fractionated on an agarose gel, then purified by phenol extraction and ethanol precipitation.

In this Example, the polynucleotide constructs are administered as naked polynucleotides via electroporation. However, the polynucleotide constructs may also be administered with transfection-facilitating agents, such as liposomes, viral sequences, viral particles, precipitating agents, etc. Such methods of delivery are known in the art.

Once the cells are transfected, homologous recombination will take place which results in the promoter being operably linked to the endogenous polynucleotide sequence. This results in the expression of polynucleotide corresponding to the polynucleotide in the cell. Expression may be detected by immunological staining, or any other method known in the art.

Fibroblasts are obtained from a subject by skin biopsy. The resulting tissue is placed in DMEM+10% fetal calf serum. Exponentially growing or early stationary phase fibroblasts are trypsinized and rinsed from the plastic surface with nutrient medium. An aliquot of the cell suspension is removed for counting, and the remaining cells are subjected to centrifugation. The supernatant is aspirated and the pellet is resuspended in 5 ml of electroporation buffer (20 mM HEPES pH 7.3, 137 mM NaCl, 5 mM KCl, 0.7 mM Na₂ HPO₄, 6 mM dextrose). The cells are recentrifuged, the supernatant aspirated, and the cells resuspended in electroporation buffer containing 1 mg/ml acetylated bovine serum albumin. The final cell suspension contains approximately 3×10⁶ cells/ml. Electroporation should be performed immediately following resuspension.

Plasmid DNA is prepared according to standard techniques. For example, to construct a plasmid for targeting to the locus corresponding to the polynucleotide of the invention, plasmid pUC18 (MBI Fermentas, Amherst, N.Y.) is digested with HindIII. The CMV promoter is amplified by PCR with an XbaI site on the 5′ end and a BamHI site on the 3′ end. Two non-coding sequences are amplified via PCR: one non-coding sequence (fragment 1) is amplified with a HindIII site at the 5′ end and an Xba site at the 3′end; the other non-coding sequence (fragment 2) is amplified with a BamHI site at the 5′end and a HindIII site at the 3′end. The CMV promoter and the fragments (1 and 2) are digested with the appropriate enzymes (CMV promoter—XbaI and BamHI; fragment 1—XbaI; fragment 2—BamHI) and ligated together. The resulting ligation product is digested with HindIII, and ligated with the HindIII-digested pUC18 plasmid.

Plasmid DNA is added to a sterile cuvette with a 0.4 cm electrode gap (Bio-Rad). The final DNA concentration is generally at least 120 μg/ml. 0.5 ml of the cell suspension (containing approximately 1.5×10⁶ cells) is then added to the cuvette, and the cell suspension and DNA solutions are gently mixed. Electroporation is performed with a Gene-Pulser apparatus (Bio-Rad). Capacitance and voltage are set at 960 μF and 250-300 V, respectively. As voltage increases, cell survival decreases, but the percentage of surviving cells that stably incorporate the introduced DNA into their genome increases dramatically. Given these parameters, a pulse time of approximately 14-20 mSec should be observed.

Electroporated cells are maintained at room temperature for approximately 5 min, and the contents of the cuvette are then gently removed with a sterile transfer pipette. The cells are added directly to 10 ml of prewarmed nutrient media (DMEM with 15% calf serum) in a 10 cm dish and incubated at 37 degree C. The following day, the media is aspirated and replaced with 10 ml of fresh media and incubated for a further 16-24 hours.

The engineered fibroblasts are then injected into the host, either alone or after having been grown to confluence on cytodex 3 microcarrier beads. The fibroblasts now produce the protein product. The fibroblasts can then be introduced into a patient as described above.

Example 18 Method of Treatment Using Gene Therapy—In Vivo

Another aspect of the present invention is using in vivo gene therapy methods to treat disorders, diseases and conditions. The gene therapy method relates to the introduction of naked nucleic acid (DNA, RNA, and antisense DNA or RNA) sequences into an animal to increase or decrease the expression of the polypeptide. The polynucleotide of the present invention may be operatively linked to (i.e., associated with) a promoter or any other genetic elements necessary for the expression of the polypeptide by the target tissue. Such gene therapy and delivery techniques and methods are known in the art, see, for example, WO90/11092, WO98/11779; U.S. Pat. Nos. 5,693,622, 5,705,151, 5,580,859; Tabata et al., Cardiovasc. Res. 35(3):470-479 (1997); Chao et al., Pharmacol. Res. 35(6):517-522 (1997); Wolff, Neuromuscul. Disord. 7(5):314-318 (1997); Schwartz et al., Gene Ther. 3(5):405-411 (1996); Tsurumi et al., Circulation 94(12):3281-3290 (1996) (incorporated herein by reference).

The polynucleotide constructs may be delivered by any method that delivers injectable materials to the cells of an animal, such as, injection into the interstitial space of tissues (heart, muscle, skin, lung, liver, intestine and the like). The polynucleotide constructs can be delivered in a pharmaceutically acceptable liquid or aqueous carrier.

The term “naked” polynucleotide, DNA or RNA, refers to sequences that are free from any delivery vehicle that acts to assist, promote, or facilitate entry into the cell, including viral sequences, viral particles, liposome formulations, lipofectin or precipitating agents and the like. However, the polynucleotides of the present invention may also be delivered in liposome formulations (such as those taught in Feigner P. L. et al. (1995) Ann. NY Acad. Sci. 772:126-139 and Abdallah B. et al. (1995) Biol. Cell 85(1):1-7) which can be prepared by methods well known to those skilled in the art.

The polynucleotide vector constructs used in the gene therapy method are preferably constructs that will not integrate into the host genome nor will they contain sequences that allow for replication. Any strong promoter known to those skilled in the art can be used for driving the expression of DNA. Unlike other gene therapy techniques, one major advantage of introducing naked nucleic acid sequences into target cells is the transitory nature of the polynucleotide synthesis in the cells. Studies have shown that non-replicating DNA sequences can be introduced into cells to provide production of the desired polypeptide for periods of up to six months.

The polynucleotide construct can be delivered to the interstitial space of tissues within an animal, including muscle, skin, brain, lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone, cartilage, pancreas, kidney, gall bladder, stomach, intestine, testis, ovary, uterus, rectum, nervous system, eye, gland, and connective tissue. Interstitial space of the tissues comprises the intercellular fluid, mucopolysaccharide matrix among the reticular fibers of organ tissues, elastic fibers in the walls of vessels or chambers, collagen fibers of fibrous tissues, or that same matrix within connective tissue ensheathing muscle cells or in the lacunae of bone. It is similarly the space occupied by the plasma of the circulation and the lymph fluid of the lymphatic channels. Delivery to the interstitial space of muscle tissue is preferred for the reasons discussed below. They may be conveniently delivered by injection into the tissues comprising these cells. They are preferably delivered to and expressed in persistent, non-dividing cells which are differentiated, although delivery and expression may be achieved in non-differentiated or less completely differentiated cells, such as, for example, stem cells of blood or skin fibroblasts. In vivo muscle cells are particularly competent in their ability to take up and express polynucleotides.

For the naked polynucleotide injection, an effective dosage amount of DNA or RNA will be in the range of from about 0.05 g/kg body weight to about 50 mg/kg body weight. Preferably the dosage will be from about 0.005 mg/kg to about 20 mg/kg and more preferably from about 0.05 mg/kg to about 5 mg/kg. Of course, as the artisan of ordinary skill will appreciate, this dosage will vary according to the tissue site of injection. The appropriate and effective dosage of nucleic acid sequence can readily be determined by those of ordinary skill in the art and may depend on the condition being treated and the route of administration. The preferred route of administration is by the parenteral route of injection into the interstitial space of tissues. However, other parenteral routes may also be used, such as, inhalation of an aerosol formulation particularly for delivery to lungs or bronchial tissues, throat or mucous membranes of the nose. In addition, naked polynucleotide constructs can be delivered to arteries during angioplasty by the catheter used in the procedure.

The dose response effects of injected polynucleotide in muscle in vivo is determined as follows. Suitable template DNA for production of mRNA coding for polypeptide of the present invention is prepared in accordance with a standard recombinant DNA methodology. The template DNA, which may be either circular or linear, is either used as naked DNA or complexed with liposomes. The quadriceps muscles of mice are then injected with various amounts of the template DNA.

Five to six week old female and male Balb/C mice are anesthetized by intraperitoneal injection with 0.3 ml of 2.5% Avertin. A 1.5 cm incision is made on the anterior thigh, and the quadriceps muscle is directly visualized. The template DNA is injected in 0.1 ml of carrier in a 1 cc syringe through a 27 gauge needle over one minute, approximately 0.5 cm from the distal insertion site of the muscle into the knee and about 0.2 cm deep. A suture is placed over the injection site for future localization, and the skin is closed with stainless steel clips.

After an appropriate incubation time (e.g., 7 days) muscle extracts are prepared by excising the entire quadriceps. Every fifth 15 um cross-section of the individual quadriceps muscles is histochemically stained for protein expression. A time course for protein expression may be done in a similar fashion except that quadriceps from different mice are harvested at different times. Persistence of DNA in muscle following injection may be determined by Southern blot analysis after preparing total cellular DNA and HIRT supernatants from injected and control mice. The results of the above experimentation in mice can be used to extrapolate proper dosages and other treatment parameters in humans and other animals using naked DNA.

Example 19 Transgenic Animals

The polypeptides of the invention can also be expressed in transgenic animals. Animals of any species, including, but not limited to, mice, rats, rabbits, hamsters, guinea pigs, pigs, micro-pigs, goats, sheep, cows and non-human primates, e.g., baboons, monkeys, and chimpanzees may be used to generate transgenic animals. In a specific embodiment, techniques described herein or otherwise known in the art, are used to express polypeptides of the invention in humans, as part of a gene therapy protocol.

Any technique known in the art may be used to introduce the transgene (i.e., polynucleotides of the invention) into animals to produce the founder lines of transgenic animals. Such techniques include, but are not limited to, pronuclear microinjection (Paterson et al., Appl. Microbiol. Biotechnol. 40:691-698 (1994); Carver et al., Biotechnology (NY) 11:1263-1270 (1993); Wright et al., Biotechnology (NY) 9:830-834 (1991); and Hoppe et al., U.S. Pat. No. 4,873,191 (1989)); retrovirus mediated gene transfer into germ lines (Van der Putten et al., Proc. Natl. Acad. Sci., USA 82:6148-6152 (1985)), blastocysts or embryos; gene targeting in embryonic stem cells (Thompson et al., Cell 56:313-321 (1989)); electroporation of cells or embryos (Lo, 1983, Mol Cell. Biol. 3:1803-1814 (1983)); introduction of the polynucleotides of the invention using a gene gun (see, e.g., Ulmer et al., Science 259:1745 (1993); introducing nucleic acid constructs into embryonic pleuripotent stem cells and transferring the stem cells back into the blastocyst; and sperm-mediated gene transfer (Lavitrano et al., Cell 57:717-723 (1989); etc. For a review of such techniques, see Gordon, “Transgenic Animals,” Intl. Rev. Cytol. 115:171-229 (1989), which is incorporated by reference herein in its entirety.

Any technique known in the art may be used to produce transgenic clones containing polynucleotides of the invention, for example, nuclear transfer into enucleated oocytes of nuclei from cultured embryonic, fetal, or adult cells induced to quiescence (Campell et al., Nature 380:64-66 (1996); Wilmut et al., Nature 385:810-813 (1997)).

The present invention provides for transgenic animals that carry the transgene in all their cells, as well as animals which carry the transgene in some, but not all their cells, i.e., mosaic animals or chimeric. The transgene may be integrated as a single transgene or as multiple copies such as in concatamers, e.g., head-to-head tandems or head-to-tail tandems. The transgene may also be selectively introduced into and activated in a particular cell type by following, for example, the teaching of Lasko et al. (Lasko et al., Proc. Natl. Acad. Sci. USA 89:6232-6236 (1992)). The regulatory sequences required for such a cell-type specific activation will depend upon the particular cell type of interest, and will be apparent to those of skill in the art. When it is desired that the polynucleotide transgene be integrated into the chromosomal site of the endogenous gene, gene targeting is preferred. Briefly, when such a technique is to be utilized, vectors containing some nucleotide sequences homologous to the endogenous gene are designed for the purpose of integrating, via homologous recombination with chromosomal sequences, into and disrupting the function of the nucleotide sequence of the endogenous gene. The transgene may also be selectively introduced into a particular cell type, thus inactivating the endogenous gene in only that cell type, by following, for example, the teaching of Gu et al. (Gu et al., Science 265:103-106 (1994)). The regulatory sequences required for such a cell-type specific inactivation will depend upon the particular cell type of interest, and will be apparent to those of skill in the art.

Once transgenic animals have been generated, the expression of the recombinant gene may be assayed utilizing standard techniques. Initial screening may be accomplished by Southern blot analysis or PCR techniques to analyze animal tissues to verify that integration of the transgene has taken place. The level of mRNA expression of the transgene in the tissues of the transgenic animals may also be assessed using techniques which include, but are not limited to, Northern blot analysis of tissue samples obtained from the animal, in situ hybridization analysis, and reverse transcriptase-PCR (rt-PCR). Samples of transgenic gene-expressing tissue may also be evaluated immunocytochemically or immunohistochemically using antibodies specific for the transgene product.

Once the founder animals are produced, they may be bred, inbred, outbred, or crossbred to produce colonies of the particular animal. Examples of such breeding strategies include, but are not limited to: outbreeding of founder animals with more than one integration site in order to establish separate lines; inbreeding of separate lines in order to produce compound transgenics that express the transgene at higher levels because of the effects of additive expression of each transgene; crossing of heterozygous transgenic animals to produce animals homozygous for a given integration site in order to both augment expression and eliminate the need for screening of animals by DNA analysis; crossing of separate homozygous lines to produce compound heterozygous or homozygous lines; and breeding to place the transgene on a distinct background that is appropriate for an experimental model of interest.

Transgenic animals of the invention have uses which include, but are not limited to, animal model systems useful in elaborating the biological function of polypeptides of the present invention, studying conditions and/or disorders associated with aberrant expression, and in screening for compounds effective in ameliorating such conditions and/or disorders.

Example 20 Knock-Out Animals

Endogenous gene expression can also be reduced by inactivating or “knocking out” the gene and/or its promoter using targeted homologous recombination. (e.g., see Smithies et al., Nature 317:230-234 (1985); Thomas & Capecchi, Cell 51:503-512 (1987); Thompson et al., Cell 5:313-321 (1989); each of which is incorporated by reference herein in its entirety). For example, a mutant, non-functional polynucleotide of the invention (or a completely unrelated DNA sequence) flanked by DNA homologous to the endogenous polynucleotide sequence (either the coding regions or regulatory regions of the gene) can be used, with or without a selectable marker and/or a negative selectable marker, to transfect cells that express polypeptides of the invention in vivo. In another embodiment, techniques known in the art are used to generate knockouts in cells that contain, but do not express the gene of interest. Insertion of the DNA construct, via targeted homologous recombination, results in inactivation of the targeted gene. Such approaches are particularly suited in research and agricultural fields where modifications to embryonic stem cells can be used to generate animal offspring with an inactive targeted gene (e.g., see Thomas & Capecchi 1987 and Thompson 1989, supra). However this approach can be routinely adapted for use in humans provided the recombinant DNA constructs are directly administered or targeted to the required site in vivo using appropriate viral vectors that will be apparent to those of skill in the art.

In further embodiments of the invention, cells that are genetically engineered to express the polypeptides of the invention, or alternatively, that are genetically engineered not to express the polypeptides of the invention (e.g., knockouts) are administered to a patient in vivo. Such cells may be obtained from the patient (i.e., animal, including human) or an MHC compatible donor and can include, but are not limited to fibroblasts, bone marrow cells, blood cells (e.g., lymphocytes), adipocytes, muscle cells, endothelial cells etc. The cells are genetically engineered in vitro using recombinant DNA techniques to introduce the coding sequence of polypeptides of the invention into the cells, or alternatively, to disrupt the coding sequence and/or endogenous regulatory sequence associated with the polypeptides of the invention, e.g., by transduction (using viral vectors, and preferably vectors that integrate the transgene into the cell genome) or transfection procedures, including, but not limited to, the use of plasmids, cosmids, YACs, naked DNA, electroporation, liposomes, etc. The coding sequence of the polypeptides of the invention can be placed under the control of a strong constitutive or inducible promoter or promoter/enhancer to achieve expression, and preferably secretion, of the polypeptides of the invention. The engineered cells which express and preferably secrete the polypeptides of the invention can be introduced into the patient systemically, e.g., in the circulation, or intraperitoneally.

Alternatively, the cells can be incorporated into a matrix and implanted in the body, e.g., genetically engineered fibroblasts can be implanted as part of a skin graft; genetically engineered endothelial cells can be implanted as part of a lymphatic or vascular graft. (See, for example, Anderson et al. U.S. Pat. No. 5,399,349; and Mulligan & Wilson, U.S. Pat. No. 5,460,959 each of which is incorporated by reference herein in its entirety).

When the cells to be administered are non-autologous or non-MHC compatible cells, they can be administered using well known techniques which prevent the development of a host immune response against the introduced cells. For example, the cells may be introduced in an encapsulated form which, while allowing for an exchange of components with the immediate extracellular environment, does not allow the introduced cells to be recognized by the host immune system.

Transgenic and “knock-out” animals of the invention have uses which include, but are not limited to, animal model systems useful in elaborating the biological function of polypeptides of the present invention, studying conditions and/or disorders associated with aberrant expression, and in screening for compounds effective in ameliorating such conditions and/or disorders.

Example 21 Assays Detecting Stimulation or Inhibition of B cell Proliferation and Differentiation

Generation of functional humoral immune responses requires both soluble and cognate signaling between B-lineage cells and their microenvironment. Signals may impart a positive stimulus that allows a B-lineage cell to continue its programmed development, or a negative stimulus that instructs the cell to arrest its current developmental pathway. To date, numerous stimulatory and inhibitory signals have been found to influence B cell responsiveness including IL-2, IL-4, IL-5, IL-6, IL-7, IL10, IL-13, IL-14 and IL-15. Interestingly, these signals are by themselves weak effectors but can, in combination with various co-stimulatory proteins, induce activation, proliferation, differentiation, homing, tolerance and death among B cell populations.

One of the best studied classes of B-cell co-stimulatory proteins is the TNF-superfamily. Within this family CD40, CD27, and CD30 along with their respective ligands CD154, CD70, and CD153 have been found to regulate a variety of immune responses. Assays which allow for the detection and/or observation of the proliferation and differentiation of these B-cell populations and their precursors are valuable tools in determining the effects various proteins may have on these B-cell populations in terms of proliferation and differentiation. Listed below are two assays designed to allow for the detection of the differentiation, proliferation, or inhibition of B-cell populations and their precursors.

In Vitro Assay—Agonists or antagonists of the invention can be assessed for its ability to induce activation, proliferation, differentiation or inhibition and/or death in B-cell populations and their precursors. The activity of the agonists or antagonists of the invention on purified human tonsillar B cells, measured qualitatively over the dose range from 0.1 to 10,000 ng/mL, is assessed in a standard B-lymphocyte co-stimulation assay in which purified tonsillar B cells are cultured in the presence of either formalin-fixed Staphylococcus aureus Cowan I (SAC) or immobilized anti-human IgM antibody as the priming agent. Second signals such as IL-2 and IL-15 synergize with SAC and IgM crosslinking to elicit B cell proliferation as measured by tritiated-thymidine incorporation. Novel synergizing agents can be readily identified using this assay. The assay involves isolating human tonsillar B cells by magnetic bead (MACS) depletion of CD3-positive cells. The resulting cell population is greater than 95% B cells as assessed by expression of CD45R(B220).

Various dilutions of each sample are placed into individual wells of a 96-well plate to which are added 10⁵ B-cells suspended in culture medium (RPMI 1640 containing 10% FBS, 5×10⁻⁵M 2ME, 100 U/ml penicillin, 10 ug/ml streptomycin, and 10⁻⁵ dilution of SAC) in a total volume of 150 ul. Proliferation or inhibition is quantitated by a 20 h pulse (1 uCi/well) with 3H-thymidine (6.7 Ci/mM) beginning 72 h post factor addition. The positive and negative controls are IL2 and medium respectively.

In vivo Assay—BALB/c mice are injected (i.p.) twice per day with buffer only, or 2 mg/Kg of agonists or antagonists of the invention, or truncated forms thereof. Mice receive this treatment for 4 consecutive days, at which time they are sacrificed and various tissues and serum collected for analyses. Comparison of H&E sections from normal spleens and spleens treated with agonists or antagonists of the invention identify the results of the activity of the agonists or antagonists on spleen cells, such as the diffusion of peri-arterial lymphatic sheaths, and/or significant increases in the nucleated cellularity of the red pulp regions, which may indicate the activation of the differentiation and proliferation of B-cell populations. Immunohistochemical studies using a B cell marker, anti-CD45R(B220), are used to determine whether any physiological changes to splenic cells, such as splenic disorganization, are due to increased B-cell representation within loosely defined B-cell zones that infiltrate established T-cell regions.

Flow cytometric analyses of the spleens from mice treated with agonist or antagonist is used to indicate whether the agonists or antagonists specifically increases the proportion of ThB+, CD45R(B220)dull B cells over that which is observed in control mice.

Likewise, a predicted consequence of increased mature B-cell representation in vivo is a relative increase in serum Ig titers. Accordingly, serum IgM and IgA levels are compared between buffer and agonists or antagonists-treated mice.

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 22 T Cell Proliferation Assay

A CD3-induced proliferation assay is performed on PBMCs and is measured by the uptake of 3H-thymidine. The assay is performed as follows. Ninety-six well plates are coated with 100 μl/well of mAb to CD3 (HIT3a, Pharmingen) or isotype-matched control mAb (B33.1) overnight at 4 degrees C. (1 μg/ml in 0.05M bicarbonate buffer, pH 9.5), then washed three times with PBS. PBMC are isolated by F/H gradient centrifugation from human peripheral blood and added to quadruplicate wells (5×10⁴/well) of mAb coated plates in RPMI containing 10% FCS and P/S in the presence of varying concentrations of agonists or antagonists of the invention (total volume 200 ul). Relevant protein buffer and medium alone are controls. After 48 hr. culture at 37 degrees C., plates are spun for 2 min. at 1000 rpm and 100 μl of supernatant is removed and stored −20 degrees C. for measurement of IL-2 (or other cytokines) if effect on proliferation is observed. Wells are supplemented with 100 ul of medium containing 0.5 uCi of ³H-thymidine and cultured at 37 degrees C. for 18-24 hr. Wells are harvested and incorporation of ³H-thymidine used as a measure of proliferation. Anti-CD3 alone is the positive control for proliferation. IL-2 (100 U/ml) is also used as a control which enhances proliferation. Control antibody which does not induce proliferation of T cells is used as the negative control for the effects of agonists or antagonists of the invention.

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 23 Effect of Agonists or Antagonists of the Invention on the Expression of MHC Class II, Costimulatory and Adhesion Molecules and Cell Differentiation of Monocytes and Monocyte-Derived Human Dendritic Cells

Dendritic cells are generated by the expansion of proliferating precursors found in the peripheral blood: adherent PBMC or elutriated monocytic fractions are cultured for 7-10 days with GM-CSF (50 ng/ml) and IL-4 (20 ng/ml). These dendritic cells have the characteristic phenotype of immature cells (expression of CD1, CD80, CD86, CD40 and MHC class II antigens). Treatment with activating factors, such as TNF-α, causes a rapid change in surface phenotype (increased expression of MHC class I and II, costimulatory and adhesion molecules, downregulation of FCγRII, upregulation of CD83). These changes correlate with increased antigen-presenting capacity and with functional maturation of the dendritic cells.

FACS analysis of surface antigens is performed as follows. Cells are treated 1-3 days with increasing concentrations of agonist or antagonist of the invention or LPS (positive control), washed with PBS containing 1% BSA and 0.02 mM sodium azide, and then incubated with 1:20 dilution of appropriate FITC- or PE-labeled monoclonal antibodies for 30 minutes at 4 degrees C. After an additional wash, the labeled cells are analyzed by flow cytometry on a FACScan (Becton Dickinson).

Effect on the production of cytokines. Cytokines generated by dendritic cells, in particular IL-12, are important in the initiation of T-cell dependent immune responses. IL-12 strongly influences the development of Th1 helper T-cell immune response, and induces cytotoxic T and NK cell function. An ELISA is used to measure the IL-12 release as follows. Dendritic cells (10⁶/ml) are treated with increasing concentrations of agonists or antagonists of the invention for 24 hours. LPS (100 ng/ml) is added to the cell culture as positive control. Supernatants from the cell cultures are then collected and analyzed for IL-12 content using commercial ELISA kit (e.g., R & D Systems (Minneapolis, Minn.)). The standard protocols provided with the kits are used.

Effect on the expression of MHC Class II, costimulatory and adhesion molecules. Three major families of cell surface antigens can be identified on monocytes: adhesion molecules, molecules involved in antigen presentation, and Fc receptor. Modulation of the expression of MHC class II antigens and other costimulatory molecules, such as B7 and ICAM-1, may result in changes in the antigen presenting capacity of monocytes and ability to induce T cell activation. Increased expression of Fc receptors may correlate with improved monocyte cytotoxic activity, cytokine release and phagocytosis.

FACS analysis is used to examine the surface antigens as follows. Monocytes are treated 1-5 days with increasing concentrations of agonists or antagonists of the invention or LPS (positive control), washed with PBS containing 1% BSA and 0.02 mM sodium azide, and then incubated with 1:20 dilution of appropriate FITC- or PE-labeled monoclonal antibodies for 30 minutes at 4 degrees C. After an additional wash, the labeled cells are analyzed by flow cytometry on a FACScan (Becton Dickinson).

Monocyte activation and/or increased survival. Assays for molecules that activate (or alternatively, inactivate) monocytes and/or increase monocyte survival (or alternatively, decrease monocyte survival) are known in the art and may routinely be applied to determine whether a molecule of the invention functions as an inhibitor or activator of monocytes. Agonists or antagonists of the invention can be screened using the three assays described below. For each of these assays, Peripheral blood mononuclear cells (PBMC) are purified from single donor leukopacks (American Red Cross, Baltimore, Md.) by centrifugation through a Histopaque gradient (Sigma). Monocytes are isolated from PBMC by counterflow centrifugal elutriation.

Monocyte Survival Assay. Human peripheral blood monocytes progressively lose viability when cultured in absence of serum or other stimuli. Their death results from internally regulated processes (apoptosis). Addition to the culture of activating factors, such as TNF-alpha dramatically improves cell survival and prevents DNA fragmentation. Propidium iodide (PI) staining is used to measure apoptosis as follows. Monocytes are cultured for 48 hours in polypropylene tubes in serum-free medium (positive control), in the presence of 100 ng/ml TNF-alpha (negative control), and in the presence of varying concentrations of the compound to be tested. Cells are suspended at a concentration of 2×10⁶/ml in PBS containing PI at a final concentration of 5 μg/ml, and then incubated at room temperature for 5 minutes before FACScan analysis. PI uptake has been demonstrated to correlate with DNA fragmentation in this experimental paradigm.

Effect on cytokine release. An important function of monocytes/macrophages is their regulatory activity on other cellular populations of the immune system through the release of cytokines after stimulation. An ELISA to measure cytokine release is performed as follows. Human monocytes are incubated at a density of 5×10⁵ cells/ml with increasing concentrations of agonists or antagonists of the invention and under the same conditions, but in the absence of agonists or antagonists. For IL-12 production, the cells are primed overnight with IFN (100 U/ml) in the presence of agonist or antagonist of the invention. LPS (10 ng/ml) is then added. Conditioned media are collected after 24 h and kept frozen until use. Measurement of TNF-alpha, IL-10, MCP-1 and IL-8 is then performed using a commercially available ELISA kit (e.g., R & D Systems (Minneapolis, Minn.)) and applying the standard protocols provided with the kit.

Oxidative burst. Purified monocytes are plated in 96-w plate at 2-1×10⁵ cell/well. Increasing concentrations of agonists or antagonists of the invention are added to the wells in a total volume of 0.2 ml culture medium (RPMI 1640+10% FCS, glutamine and antibiotics). After 3 days incubation, the plates are centrifuged and the medium is removed from the wells. To the macrophage monolayers, 0.2 ml per well of phenol red solution (140 mM NaCl, 10 mM potassium phosphate buffer pH 7.0, 5.5 mM dextrose, 0.56 mM phenol red and 19 U/ml of HRPO) is added, together with the stimulant (200 nM PMA). The plates are incubated at 37° C. for 2 hours and the reaction is stopped by adding 20 μl 1N NaOH per well. The absorbance is read at 610 nm. To calculate the amount of H₂O₂ produced by the macrophages, a standard curve of a H₂O₂ solution of known molarity is performed for each experiment.

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 24 Biological Effects of Agonists or Antagonists of the Invention

Astrocyte and Neuronal Assays

Agonists or antagonists of the invention, expressed in Escherichia coli and purified as described above, can be tested for activity in promoting the survival, neurite outgrowth, or phenotypic differentiation of cortical neuronal cells and for inducing the proliferation of glial fibrillary acidic protein immunopositive cells, astrocytes. The selection of cortical cells for the bioassay is based on the prevalent expression of FGF-1 and FGF-2 in cortical structures and on the previously reported enhancement of cortical neuronal survival resulting from FGF-2 treatment. A thymidine incorporation assay, for example, can be used to elucidate an agonist or antagonist of the invention's activity on these cells.

Moreover, previous reports describing the biological effects of FGF-2 (basic FGF) on cortical or hippocampal neurons in vitro have demonstrated increases in both neuron survival and neurite outgrowth (Walicke et al., “Fibroblast growth factor promotes survival of dissociated hippocampal neurons and enhances neurite extension.” Proc. Natl. Acad. Sci. USA 83:3012-3016. (1986), assay herein incorporated by reference in its entirety). However, reports from experiments done on PC-12 cells suggest that these two responses are not necessarily synonymous and may depend on not only which FGF is being tested but also on which receptor(s) are expressed on the target cells. Using the primary cortical neuronal culture paradigm, the ability of an agonist or antagonist of the invention to induce neurite outgrowth can be compared to the response achieved with FGF-2 using, for example, a thymidine incorporation assay.

Fibroblast and Endothelial Cell Assays

Human lung fibroblasts are obtained from Clonetics (San Diego, Calif.) and maintained in growth media from Clonetics. Dermal microvascular endothelial cells are obtained from Cell Applications (San Diego, Calif.). For proliferation assays, the human lung fibroblasts and dermal microvascular endothelial cells can be cultured at 5,000 cells/well in a 96-well plate for one day in growth medium. The cells are then incubated for one day in 0.1% BSA basal medium. After replacing the medium with fresh 0.1% BSA medium, the cells are incubated with the test proteins for 3 days. Alamar Blue (Alamar Biosciences, Sacramento, Calif.) is added to each well to a final concentration of 10%. The cells are incubated for 4 hr. Cell viability is measured by reading in a CytoFluor fluorescence reader. For the PGE₂ assays, the human lung fibroblasts are cultured at 5,000 cells/well in a 96-well plate for one day. After a medium change to 0.1% BSA basal medium, the cells are incubated with FGF-2 or agonists or antagonists of the invention with or without IL-1α for 24 hours. The supernatants are collected and assayed for PGE₂ by EIA kit (Cayman, Ann Arbor, Mich.). For the IL-6 assays, the human lung fibroblasts are cultured at 5,000 cells/well in a 96-well plate for one day. After a medium change to 0.1% BSA basal medium, the cells are incubated with FGF-2 or with or without agonists or antagonists of the invention IL-1α for 24 hours. The supernatants are collected and assayed for IL-6 by ELISA kit (Endogen, Cambridge, Mass.).

Human lung fibroblasts are cultured with FGF-2 or agonists or antagonists of the invention for 3 days in basal medium before the addition of Alamar Blue to assess effects on growth of the fibroblasts. FGF-2 should show a stimulation at 10⁻²⁵⁰⁰ ng/ml which can be used to compare stimulation with agonists or antagonists of the invention.

Parkinson Models.

The loss of motor function in Parkinson's disease is attributed to a deficiency of striatal dopamine resulting from the degeneration of the nigrostriatal dopaminergic projection neurons. An animal model for Parkinson's that has been extensively characterized involves the systemic administration of 1-methyl-4 phenyl 1,2,3,6-tetrahydropyridine (MPTP). In the CNS, MPTP is taken-up by astrocytes and catabolized by monoamine oxidase B to 1-methyl-4-phenyl pyridine (MPP⁺) and released. Subsequently, MPP⁺ is actively accumulated in dopaminergic neurons by the high-affinity reuptake transporter for dopamine. MPP⁺ is then concentrated in mitochondria by the electrochemical gradient and selectively inhibits nicotidamide adenine disphosphate: ubiquinone oxidoreductionase (complex I), thereby interfering with electron transport and eventually generating oxygen radicals.

It has been demonstrated in tissue culture paradigms that FGF-2 (basic FGF) has trophic activity towards nigral dopaminergic neurons (Ferrari et al., Dev. Biol. 1989). Recently, Dr. Unsicker's group has demonstrated that administering FGF-2 in gel foam implants in the striatum results in the near complete protection of nigral dopaminergic neurons from the toxicity associated with MPTP exposure (Otto and Unsicker, J. Neuroscience, 1990).

Based on the data with FGF-2, agonists or antagonists of the invention can be evaluated to determine whether it has an action similar to that of FGF-2 in enhancing dopaminergic neuronal survival in vitro and it can also be tested in vivo for protection of dopaminergic neurons in the striatum from the damage associated with MPTP treatment. The potential effect of an agonist or antagonist of the invention is first examined in vitro in a dopaminergic neuronal cell culture paradigm. The cultures are prepared by dissecting the midbrain floor plate from gestation day 14 Wistar rat embryos. The tissue is dissociated with trypsin and seeded at a density of 200,000 cells/cm² on polyorthinine-laminin coated glass coverslips. The cells are maintained in Dulbecco's Modified Eagle's medium and F12 medium containing hormonal supplements (N1). The cultures are fixed with paraformaldehyde after 8 days in vitro and are processed for tyrosine hydroxylase, a specific marker for dopaminergic neurons, immunohistochemical staining. Dissociated cell cultures are prepared from embryonic rats. The culture medium is changed every third day and the factors are also added at that time.

Since the dopaminergic neurons are isolated from animals at gestation day 14, a developmental time which is past the stage when the dopaminergic precursor cells are proliferating, an increase in the number of tyrosine hydroxylase immunopositive neurons would represent an increase in the number of dopaminergic neurons surviving in vitro. Therefore, if an agonist or antagonist of the invention acts to prolong the survival of dopaminergic neurons, it would suggest that the agonist or antagonist may be involved in Parkinson's Disease.

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 25 The Effect of Agonists or Antagonists of the Invention on the Growth of Vascular Endothelial Cells

On day 1, human umbilical vein endothelial cells (HUVEC) are seeded at 2-5×10⁴ cells/35 mm dish density in M199 medium containing 4% fetal bovine serum (FBS), 16 units/ml heparin, and 50 units/ml endothelial cell growth supplements (ECGS, Biotechnique, Inc.). On day 2, the medium is replaced with M199 containing 10% FBS, 8 units/ml heparin. An agonist or antagonist of the invention, and positive controls, such as VEGF and basic FGF (bFGF) are added, at varying concentrations. On days 4 and 6, the medium is replaced. On day 8, cell number is determined with a Coulter Counter.

An increase in the number of HUVEC cells indicates that the compound of the invention may proliferate vascular endothelial cells, while a decrease in the number of HUVEC cells indicates that the compound of the invention inhibits vascular endothelial cells.

The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.

Example 26 Rat Corneal Wound Healing Model

This animal model shows the effect of an agonist or antagonist of the invention on neovascularization. The experimental protocol includes:

a) Making a 1-1.5 mm long incision from the center of cornea into the stromal layer.

b) Inserting a spatula below the lip of the incision facing the outer corner of the eye.

c) Making a pocket (its base is 1-1.5 mm form the edge of the eye).

d) Positioning a pellet, containing 50 ng-5 ug of an agonist or antagonist of the invention, within the pocket.

e) Treatment with an agonist or antagonist of the invention can also be applied topically to the corneal wounds in a dosage range of 20 mg-500 mg (daily treatment for five days).

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 27 Diabetic Mouse and Glucocorticoid-Impaired Wound Heating Models

Diabetic db+/db+ Mouse Model.

To demonstrate that an agonist or antagonist of the invention accelerates the healing process, the genetically diabetic mouse model of wound healing is used. The full thickness wound healing model in the db+/db+ mouse is a well characterized, clinically relevant and reproducible model of impaired wound healing. Healing of the diabetic wound is dependent on formation of granulation tissue and re-epithelialization rather than contraction (Gartner, M. H. et al., J. Surg. Res. 52:389 (1992); Greenhalgh, D. G. et al., Am. J. Pathol. 136:1235 (1990)).

The diabetic animals have many of the characteristic features observed in Type II diabetes mellitus. Homozygous (db+/db+) mice are obese in comparison to their normal heterozygous (db+/+m) littermates. Mutant diabetic (db+/db+) mice have a single autosomal recessive mutation on chromosome 4 (db+) (Coleman et al. Proc. Natl. Acad. Sci. USA 77:283-293 (1982)). Animals show polyphagia, polydipsia and polyuria. Mutant diabetic mice (db+/db+) have elevated blood glucose, increased or normal insulin levels, and suppressed cell-mediated immunity (Mandel et al., J. Immunol. 120:1375 (1978); Debray-Sachs, M. et al., Clin. Exp. Immunol. 51(1):1-7 (1983); Leiter et al., Am. J. of Pathol 114:46-55 (1985)). Peripheral neuropathy, myocardial complications, and microvascular lesions, basement membrane thickening and glomerular filtration abnormalities have been described in these animals (Norido, F. et al., Exp. Neurol. 83(2):221-232 (1984); Robertson et al., Diabetes 29(1):60-67 (1980); Giacomelli et al., Lab Invest. 40(4):460-473 (1979); Coleman, D. L., ^(Diabetes) 31 (Suppl):1-6 (1982)). These homozygous diabetic mice develop hyperglycemia that is resistant to insulin analogous to human type II diabetes (Mandel et al., J. Immunol. 120:1375-1377 (1978)).

The characteristics observed in these animals suggests that healing in this model may be similar to the healing observed in human diabetes (Greenhalgh, et al., Am. J. of Pathol. 136:1235-1246 (1990)).

Genetically diabetic female C57BL/KsJ (db+/db+) mice and their non-diabetic (db+/+m) heterozygous littermates are used in this study (Jackson Laboratories). The animals are purchased at 6 weeks of age and are 8 weeks old at the beginning of the study. Animals are individually housed and received food and water ad libitum. All manipulations are performed using aseptic techniques. The experiments are conducted according to the rules and guidelines of Human Genome Sciences, Inc. Institutional Animal Care and Use Committee and the Guidelines for the Care and Use of Laboratory Animals.

Wounding protocol is performed according to previously reported methods (Tsuboi, R. and Rifkin, D. B., J. Exp. Med. 172:245-251 (1990)). Briefly, on the day of wounding, animals are anesthetized with an intraperitoneal injection of Avertin (0.01 mg/mL), 2,2,2-tribromoethanol and 2-methyl-2-butanol dissolved in deionized water. The dorsal region of the animal is shaved and the skin washed with 70% ethanol solution and iodine. The surgical area is dried with sterile gauze prior to wounding. An 8 mm full-thickness wound is then created using a Keyes tissue punch. Immediately following wounding, the surrounding skin is gently stretched to eliminate wound expansion. The wounds are left open for the duration of the experiment. Application of the treatment is given topically for 5 consecutive days commencing on the day of wounding. Prior to treatment, wounds are gently cleansed with sterile saline and gauze sponges.

Wounds are visually examined and photographed at a fixed distance at the day of surgery and at two day intervals thereafter. Wound closure is determined by daily measurement on days 1-5 and on day 8. Wounds are measured horizontally and vertically using a calibrated Jameson caliper. Wounds are considered healed if granulation tissue is no longer visible and the wound is covered by a continuous epithelium.

An agonist or antagonist of the invention is administered using at a range different doses, from 4 mg to 500 mg per wound per day for 8 days in vehicle. Vehicle control groups received 50 mL of vehicle solution.

Animals are euthanized on day 8 with an intraperitoneal injection of sodium pentobarbital (300 mg/kg). The wounds and surrounding skin are then harvested for histology and immunohistochemistry. Tissue specimens are placed in 10% neutral buffered formalin in tissue cassettes between biopsy sponges for further processing.

Three groups of 10 animals each (5 diabetic and 5 non-diabetic controls) are evaluated: 1) Vehicle placebo control, 2) untreated group, and 3) treated group.

Wound closure is analyzed by measuring the area in the vertical and horizontal axis and obtaining the total square area of the wound. Contraction is then estimated by establishing the differences between the initial wound area (day 0) and that of post treatment (day 8). The wound area on day 1 is 64 mm², the corresponding size of the dermal punch. Calculations are made using the following formula: [Open area on day 8]−[Open area on day 1]/[Open area on day 1]

Specimens are fixed in 10% buffered formalin and paraffin embedded blocks are sectioned perpendicular to the wound surface (5 mm) and cut using a Reichert-Jung microtome. Routine hematoxylin-eosin (H&E) staining is performed on cross-sections of bisected wounds. Histologic examination of the wounds are used to assess whether the healing process and the morphologic appearance of the repaired skin is altered by treatment with an agonist or antagonist of the invention. This assessment included verification of the presence of cell accumulation, inflammatory cells, capillaries, fibroblasts, re-epithelialization and epidermal maturity (Greenhalgh, D. G. et al., Am. J. Pathol. 136:1235 (1990)). A calibrated lens micrometer is used by a blinded observer.

Tissue sections are also stained immunohistochemically with a polyclonal rabbit anti-human keratin antibody using ABC Elite detection system. Human skin is used as a positive tissue control while non-immune IgG is used as a negative control. Keratinocyte growth is determined by evaluating the extent of reepithelialization of the wound using a calibrated lens micrometer.

Proliferating cell nuclear antigen/cyclin (PCNA) in skin specimens is demonstrated by using anti-PCNA antibody (1:50) with an ABC Elite detection system. Human colon cancer served as a positive tissue control and human brain tissue is used as a negative tissue control. Each specimen included a section with omission of the primary antibody and substitution with non-immune mouse IgG. Ranking of these sections is based on the extent of proliferation on a scale of 0-8, the lower side of the scale reflecting slight proliferation to the higher side reflecting intense proliferation.

Experimental data are analyzed using an unpaired t test. A p value of <0.05 is considered significant.

Steroid Impaired Rat Model

The inhibition of wound healing by steroids has been well documented in various in vitro and in vivo systems (Wahl, Glucocorticoids and Wound healing. In: Anti-Inflammatory Steroid Action: Basic and Clinical Aspects. 280-302 (1989); Wahl et al., J. Immunol. 115: 476-481 (1975); Werb et al., J. Exp. Med. 147:1684-1694 (1978)). Glucocorticoids retard wound healing by inhibiting angiogenesis, decreasing vascular permeability (Ebert et al., An. Intern. Med. 37:701-705 (1952)), fibroblast proliferation, and collagen synthesis (Beck et al., Growth Factors. 5: 295-304 (1991); Haynes et al., J. Clin. Invest. 61: 703-797 (1978)) and producing a transient reduction of circulating monocytes (Haynes et al., J. Clin. Invest. 61: 703-797 (1978); Wahl, “Glucocorticoids and wound healing”, In: Antiinflammatory Steroid Action: Basic and Clinical Aspects, Academic Press, New York, pp. 280-302 (1989)). The systemic administration of steroids to impaired wound healing is a well establish phenomenon in rats (Beck et al., Growth Factors. 5: 295-304 (1991); Haynes et al., J. Clin. Invest. 61: 703-797 (1978); Wahl, “Glucocorticoids and wound healing”, In: Antiinflammatory Steroid Action: Basic and Clinical Aspects, Academic Press, New York, pp. 280-302 (1989); Pierce et al., Proc. Natl. Acad. Sci. USA 86: 2229-2233 (1989)).

To demonstrate that an agonist or antagonist of the invention can accelerate the healing process, the effects of multiple topical applications of the agonist or antagonist on full thickness excisional skin wounds in rats in which healing has been impaired by the systemic administration of methylprednisolone is assessed.

Young adult male Sprague Dawley rats weighing 250-300 g (Charles River Laboratories) are used in this example. The animals are purchased at 8 weeks of age and are 9 weeks old at the beginning of the study. The healing response of rats is impaired by the systemic administration of methylprednisolone (17 mg/kg/rat intramuscularly) at the time of wounding. Animals are individually housed and received food and water ad libitum. All manipulations are performed using aseptic techniques. This study is conducted according to the rules and guidelines of Human Genome Sciences, Inc. Institutional Animal Care and Use Committee and the Guidelines for the Care and Use of Laboratory Animals.

The wounding protocol is followed according to section A, above. On the day of wounding, animals are anesthetized with an intramuscular injection of ketamine (50 mg/kg) and xylazine (5 mg/kg). The dorsal region of the animal is shaved and the skin washed with 70% ethanol and iodine solutions. The surgical area is dried with sterile gauze prior to wounding. An 8 mm full-thickness wound is created using a Keyes tissue punch. The wounds are left open for the duration of the experiment. Applications of the testing materials are given topically once a day for 7 consecutive days commencing on the day of wounding and subsequent to methylprednisolone administration. Prior to treatment, wounds are gently cleansed with sterile saline and gauze sponges.

Wounds are visually examined and photographed at a fixed distance at the day of wounding and at the end of treatment. Wound closure is determined by daily measurement on days 1-5 and on day 8. Wounds are measured horizontally and vertically using a calibrated Jameson caliper. Wounds are considered healed if granulation tissue is no longer visible and the wound is covered by a continuous epithelium.

The agonist or antagonist of the invention is administered using at a range different doses, from 4 mg to 500 mg per wound per day for 8 days in vehicle. Vehicle control groups received 50 mL of vehicle solution.

Animals are euthanized on day 8 with an intraperitoneal injection of sodium pentobarbital (300 mg/kg). The wounds and surrounding skin are then harvested for histology. Tissue specimens are placed in 10% neutral buffered formalin in tissue cassettes between biopsy sponges for further processing.

Three groups of 10 animals each (5 with methylprednisolone and 5 without glucocorticoid) are evaluated: 1) Untreated group 2) Vehicle placebo control 3) treated groups.

Wound closure is analyzed by measuring the area in the vertical and horizontal axis and obtaining the total area of the wound. Closure is then estimated by establishing the differences between the initial wound area (day 0) and that of post treatment (day 8). The wound area on day 1 is 64 mm², the corresponding size of the dermal punch. Calculations are made using the following formula: [Open area on day 8]−[Open area on day 1]/[Open area on day 1]

Specimens are fixed in 10% buffered formalin and paraffin embedded blocks are sectioned perpendicular to the wound surface (5 mm) and cut using an Olympus microtome. Routine hematoxylin-eosin (H&E) staining is performed on cross-sections of bisected wounds. Histologic examination of the wounds allows assessment of whether the healing process and the morphologic appearance of the repaired skin is improved by treatment with an agonist or antagonist of the invention. A calibrated lens micrometer is used by a blinded observer to determine the distance of the wound gap.

Experimental data are analyzed using an unpaired t test. A p value of <0.05 is considered significant.

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 28 Lymphadema Animal Model

The purpose of this experimental approach is to create an appropriate and consistent lymphedema model for testing the therapeutic effects of an agonist or antagonist of the invention in lymphangiogenesis and re-establishment of the lymphatic circulatory system in the rat hind limb. Effectiveness is measured by swelling volume of the affected limb, quantification of the amount of lymphatic vasculature, total blood plasma protein, and histopathology. Acute lymphedema is observed for 7-10 days. Perhaps more importantly, the chronic progress of the edema is followed for up to 3-4 weeks.

Prior to beginning surgery, blood sample is drawn for protein concentration analysis. Male rats weighing approximately ˜350 g are dosed with Pentobarbital. Subsequently, the right legs are shaved from knee to hip. The shaved area is swabbed with gauze soaked in 70% EtOH. Blood is drawn for serum total protein testing. Circumference and volumetric measurements are made prior to injecting dye into paws after marking 2 measurement levels (0.5 cm above heel, at mid-pt of dorsal paw). The intradermal dorsum of both right and left paws are injected with 0.05 ml of 1% Evan's Blue. Circumference and volumetric measurements are then made following injection of dye into paws.

Using the knee joint as a landmark, a mid-leg inguinal incision is made circumferentially allowing the femoral vessels to be located. Forceps and hemostats are used to dissect and separate the skin flaps. After locating the femoral vessels, the lymphatic vessel that runs along side and underneath the vessel(s) is located. The main lymphatic vessels in this area are then electrically coagulated or suture ligated.

Using a microscope, muscles in back of the leg (near the semitendinosis and adductors) are bluntly dissected. The popliteal lymph node is then located. The 2 proximal and 2 distal lymphatic vessels and distal blood supply of the popliteal node are then ligated by suturing. The popliteal lymph node, and any accompanying adipose tissue, is then removed by cutting connective tissues.

Care is taken to control any mild bleeding resulting from this procedure. After lymphatics are occluded, the skin flaps are sealed by using liquid skin (Vetbond) (AJ Buck). The separated skin edges are sealed to the underlying muscle tissue while leaving a gap of ˜0.5 cm around the leg. Skin also may be anchored by suturing to underlying muscle when necessary.

To avoid infection, animals are housed individually with mesh (no bedding). Recovering animals are checked daily through the optimal edematous peak, which typically occurred by day 5-7. The plateau edematous peak are then observed. To evaluate the intensity of the lymphedema, the circumference and volumes of 2 designated places on each paw before operation and daily for 7 days are measured. The effect of plasma proteins on lymphedema is determined and whether protein analysis is a useful testing perimeter is also investigated. The weights of both control and edematous limbs are evaluated at 2 places. Analysis is performed in a blind manner.

Circumference Measurements: Under brief gas anesthetic to prevent limb movement, a cloth tape is used to measure limb circumference. Measurements are done at the ankle bone and dorsal paw by 2 different people and those 2 readings are averaged. Readings are taken from both control and edematous limbs.

Volumetric Measurements: On the day of surgery, animals are anesthetized with Pentobarbital and are tested prior to surgery. For daily volumetrics animals are under brief halothane anesthetic (rapid immobilization and quick recovery), and both legs are shaved and equally marked using waterproof marker on legs. Legs are first dipped in water, then dipped into instrument to each marked level then measured by Buxco edema software (Chen/Victor). Data is recorded by one person, while the other is dipping the limb to marked area.

Blood-plasma protein measurements: Blood is drawn, spun, and serum separated prior to surgery and then at conclusion for total protein and Ca2⁺ comparison.

Limb Weight Comparison: After drawing blood, the animal is prepared for tissue collection. The limbs are amputated using a quillitine, then both experimental and control legs are cut at the ligature and weighed. A second weighing is done as the tibio-cacaneal joint is disarticulated and the foot is weighed.

Histological Preparations: The transverse muscle located behind the knee (popliteal) area is dissected and arranged in a metal mold, filled with freeze Gel, dipped into cold methylbutane, placed into labeled sample bags at −80EC until sectioning. Upon sectioning, the muscle is observed under fluorescent microscopy for lymphatics.

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 29 Suppression of TNF Alpha-Induced Adhesion Molecule Expression by an Agonist or Antagonist of the Invention

The recruitment of lymphocytes to areas of inflammation and angiogenesis involves specific receptor-ligand interactions between cell surface adhesion molecules (CAMs) on lymphocytes and the vascular endothelium. The adhesion process, in both normal and pathological settings, follows a multi-step cascade that involves intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial leukocyte adhesion molecule-1 (E-selectin) expression on endothelial cells (EC). The expression of these molecules and others on the vascular endothelium determines the efficiency with which leukocytes may adhere to the local vasculature and extravasate into the local tissue during the development of an inflammatory response. The local concentration of cytokines and growth factor participate in the modulation of the expression of these CAMs.

Tumor necrosis factor alpha (TNF-a), a potent proinflammatory cytokine, is a stimulator of all three CAMs on endothelial cells and may be involved in a wide variety of inflammatory responses, often resulting in a pathological outcome.

The potential of an agonist or antagonist of the invention to mediate a suppression of TNF-a induced CAM expression can be examined. A modified ELISA assay which uses ECs as a solid phase absorbent is employed to measure the amount of CAM expression on TNF-a treated ECs when co-stimulated with a member of the FGF family of proteins.

To perform the experiment, human umbilical vein endothelial cell (HUVEC) cultures are obtained from pooled cord harvests and maintained in growth medium (EGM-2; Clonetics, San Diego, Calif.) supplemented with 10% FCS and 1% penicillin/streptomycin in a 37 degree C. humidified incubator containing 5% CO₂. HUVECs are seeded in 96-well plates at concentrations of 1×10⁴ cells/well in EGM medium at 37 degree C. for 18-24 hrs or until confluent. The monolayers are subsequently washed 3 times with a serum-free solution of RPMI-1640 supplemented with 100 U/ml penicillin and 100 mg/ml streptomycin, and treated with a given cytokine and/or growth factor(s) for 24 h at 37 degree C. Following incubation, the cells are then evaluated for CAM expression.

Human Umbilical Vein Endothelial cells (HUVECs) are grown in a standard 96 well plate to confluence. Growth medium is removed from the cells and replaced with 90 ul of 199 Medium (10% FBS). Samples for testing and positive or negative controls are added to the plate in triplicate (in 10 ul volumes). Plates are incubated at 37 degree C. for either 5 h (selectin and integrin expression) or 24 h (integrin expression only). Plates are aspirated to remove medium and 100 μl of 0.1% paraformaldehyde-PBS (with Ca++ and Mg++) is added to each well. Plates are held at 4° C. for 30 min.

Fixative is then removed from the wells and wells are washed 1× with PBS(+Ca,Mg)+0.5% BSA and drained. Do not allow the wells to dry. Add 10 μl of diluted primary antibody to the test and control wells. Anti-ICAM-1-Biotin, Anti-VCAM-1-Biotin and Anti-E-selectin-Biotin are used at a concentration of 10 μg/ml (1:10 dilution of 0.1 mg/ml stock antibody). Cells are incubated at 37° C. for 30 min. in a humidified environment. Wells are washed X3 with PBS(+Ca,Mg)+0.5% BSA.

Then add 20 μl of diluted ExtrAvidin-Alkaline Phosphotase (1:5,000 dilution) to each well and incubated at 37° C. for 30 min. Wells are washed X3 with PBS(+Ca,Mg)+0.5% BSA. 1 tablet of p-Nitrophenol Phosphate pNPP is dissolved in 5 ml of glycine buffer (pH 10.4). 100 μl of pNPP substrate in glycine buffer is added to each test well. Standard wells in triplicate are prepared from the working dilution of the ExtrAvidin-Alkaline Phosphotase in glycine buffer: 1:5,000 (10⁰)>10^(−0.5)>10⁻¹>10^(−1.5)0.5 μl of each dilution is added to triplicate wells and the resulting AP content in each well is 5.50 ng, 1.74 ng, 0.55 ng, 0.18 ng. 100 μl of pNNP reagent must then be added to each of the standard wells. The plate must be incubated at 37° C. for 4 h. A volume of 50 μl of 3M NaOH is added to all wells. The results are quantified on a plate reader at 405 nm. The background subtraction option is used on blank wells filled with glycine buffer only. The template is set up to indicate the concentration of AP-conjugate in each standard well [5.50 ng; 1.74 ng; 0.55 ng; 0.18 ng]. Results are indicated as amount of bound AP-conjugate in each sample.

The studies described in this example tested activity of agonists or antagonists of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides or polypeptides of the invention (e.g., gene therapy).

Example 30 Production of Polypeptide of the Invention for High-Throughput Screening Assays

The following protocol produces a supernatant containing polypeptide of the present invention to be tested. This supernatant can then be used in the Screening Assays described in Examples 32-41.

First, dilute Poly-D-Lysine (644 587 Boehringer-Mannheim) stock solution (1 mg/ml in PBS) 1:20 in PBS (w/o calcium or magnesium 17-516F Biowhittaker) for a working solution of 50 ug/ml. Add 200 ul of this solution to each well (24 well plates) and incubate at RT for 20 minutes. Be sure to distribute the solution over each well (note: a 12-channel pipetter may be used with tips on every other channel). Aspirate off the Poly-D-Lysine solution and rinse with 1 ml PBS (Phosphate Buffered Saline). The PBS should remain in the well until just prior to plating the cells and plates may be poly-lysine coated in advance for up to two weeks.

Plate 293T cells (do not carry cells past P+20) at 2×10⁵ cells/well in 0.5 ml DMEM(Dulbecco's Modified Eagle Medium)(with 4.5 G/L glucose and L-glutamine (12-604F Biowhittaker))/10% heat inactivated FBS(14-503F Biowhittaker)/1× Penstrep(17-602E Biowhittaker). Let the cells grow overnight.

The next day, mix together in a sterile solution basin: 300 ul Lipofectamine (18324-012 Gibco/BRL) and 5 ml Optimem I (31985070 Gibco/BRL)/96-well plate. With a small volume multi-channel pipetter, aliquot approximately 2 ug of an expression vector containing a polynucleotide insert, produced by the methods described in Examples 8-10, into an appropriately labeled 96-well round bottom plate. With a multi-channel pipetter, add 50 ul of the Lipofectamine/Optimem I mixture to each well. Pipette up and down gently to mix. Incubate at RT 15-45 minutes. After about 20 minutes, use a multi-channel pipetter to add 150 ul Optimem I to each well. As a control, one plate of vector DNA lacking an insert should be transfected with each set of transfections.

Preferably, the transfection should be performed by tag-teaming the following tasks. By tag-teaming, hands on time is cut in half, and the cells do not spend too much time on PBS. First, person A aspirates off the media from four 24-well plates of cells, and then person B rinses each well with 0.5-1 ml PBS. Person A then aspirates off PBS rinse, and person B, using a12-channel pipetter with tips on every other channel, adds the 200 ul of DNA/Lipofectamine/Optimem I complex to the odd wells first, then to the even wells, to each row on the 24-well plates. Incubate at 37 degree C. for 6 hours.

While cells are incubating, prepare appropriate media, either 1% BSA in DMEM with 1× penstrep, or HGS CHO-5 media (116.6 mg/L of CaCl2 (anhyd); 0.00130 mg/L CuSO₄-5H₂O; 0.050 mg/L of Fe(NO₃)₃-9H₂O; 0.417 mg/L of FeSO₄-7H₂O; 311.80 mg/L of Kcl; 28.64 mg/L of MgCl₂; 48.84 mg/L of MgSO₄; 6995.50 mg/L of NaCl; 2400.0 mg/L of NaHCO₃; 62.50 mg/L of NaH₂PO₄-H₂0; 71.02 mg/L of Na₂HPO4; 0.4320 mg/L of ZnSO₄-7H₂O; 0.002 mg/L of Arachidonic Acid; 1.022 mg/L of Cholesterol; 0.070 mg/L of DL-alpha-Tocopherol-Acetate; 0.0520 mg/L of Linoleic Acid; 0.010 mg/L of Linolenic Acid; 0.010 mg/L of Myristic Acid; 0.010 mg/L of Oleic Acid; 0.010 mg/L of Palmitric Acid; 0.010 mg/L of Palmitic Acid; 100 mg/L of Pluronic F-68; 0.010 mg/L of Stearic Acid; 2.20 mg/L of Tween 80; 4551 mg/L of D-Glucose; 130.85 mg/ml of L-Alanine; 147.50 mg/ml of L-Arginine-HCL; 7.50 mg/ml of L-Asparagine-H₂0; 6.65 mg/ml of L-Aspartic Acid; 29.56 mg/ml of L-Cystine-2HCL-H₂0; 31.29 mg/ml of L-Cystine-2HCL; 7.35 mg/ml of L-Glutamic Acid; 365.0 mg/ml of L-Glutamine; 18.75 mg/ml of Glycine; 52.48 mg/ml of L-Histidine-HCL-H₂0; 106.97 mg/ml of L-Isoleucine; 111.45 mg/ml of L-Leucine; 163.75 mg/ml of L-Lysine HCL; 32.34 mg/ml of L-Methionine; 68.48 mg/ml of L-Phenylalainine; 40.0 mg/ml of L-Proline; 26.25 mg/ml of L-Serine; 101.05 mg/ml of L-Threonine; 19.22 mg/ml of L-Tryptophan; 91.79 mg/ml of L-Tryrosine-2Na-2H₂0; and 99.65 mg/ml of L-Valine; 0.0035 mg/L of Biotin; 3.24 mg/L of D-Ca Pantothenate; 11.78 mg/L of Choline Chloride; 4.65 mg/L of Folic Acid; 15.60 mg/L of i-Inositol; 3.02 mg/L of Niacinamide; 3.00 mg/L of Pyridoxal HCL; 0.031 mg/L of Pyridoxine HCL; 0.319 mg/L of Riboflavin; 3.17 mg/L of Thiamine HCL; 0.365 mg/L of Thymidine; 0.680 mg/L of Vitamin B₁₂; 25 mM of HEPES Buffer; 2.39 mg/L of Na Hypoxanthine; 0.105 mg/L of Lipoic Acid; 0.081 mg/L of Sodium Putrescine-2HCL; 55.0 mg/L of Sodium Pyruvate; 0.0067 mg/L of Sodium Selenite; 20 uM of Ethanolamine; 0.122 mg/L of Ferric Citrate; 41.70 mg/L of Methyl-B-Cyclodextrin complexed with Linoleic Acid; 33.33 mg/L of Methyl-B-Cyclodextrin complexed with Oleic Acid; 10 mg/L of Methyl-B-Cyclodextrin complexed with Retinal Acetate. Adjust osmolarity to 327 mOsm) with 2 mm glutamine and 1× penstrep. (BSA (81-068-3 Bayer) 100 gm dissolved in IL DMEM for a 10% BSA stock solution). Filter the media and collect 50 ul for endotoxin assay in 15 ml polystyrene conical.

The transfection reaction is terminated, preferably by tag-teaming, at the end of the incubation period. Person A aspirates off the transfection media, while person B adds 1.5 ml appropriate media to each well. Incubate at 37 degree C. for 45 or 72 hours depending on the media used: 1% BSA for 45 hours or CHO-5 for 72 hours.

On day four, using a 300 ul multichannel pipetter, aliquot 600 ul in one 1 ml deep well plate and the remaining supernatant into a 2 ml deep well. The supernatants from each well can then be used in the assays described in Examples 32-39.

It is specifically understood that when activity is obtained in any of the assays described below using a supernatant, the activity originates from either the polypeptide of the present invention directly (e.g., as a secreted protein) or by polypeptide of the present invention inducing expression of other proteins, which are then secreted into the supernatant. Thus, the invention further provides a method of identifying the protein in the supernatant characterized by an activity in a particular assay.

Example 31 Construction of GAS Reporter Construct

One signal transduction pathway involved in the differentiation and proliferation of cells is called the Jaks-STATs pathway. Activated proteins in the Jaks-STATs pathway bind to gamma activation site “GAS” elements or interferon-sensitive responsive element (“ISRE”), located in the promoter of many genes. The binding of a protein to these elements alter the expression of the associated gene.

GAS and ISRE elements are recognized by a class of transcription factors called Signal Transducers and Activators of Transcription, or “STATs.” There are six members of the STATs family. Stat1 and Stat3 are present in many cell types, as is Stat2 (as response to IFN-alpha is widespread). Stat4 is more restricted and is not in many cell types though it has been found in T helper class I, cells after treatment with IL-12. Stat5 was originally called mammary growth factor, but has been found at higher concentrations in other cells including myeloid cells. It can be activated in tissue culture cells by many cytokines.

The STATs are activated to translocate from the cytoplasm to the nucleus upon tyrosine phosphorylation by a set of kinases known as the Janus Kinase (“Jaks”) family. Jaks represent a distinct family of soluble tyrosine kinases and include Tyk2, Jak1, Jak2, and Jak3. These kinases display significant sequence similarity and are generally catalytically inactive in resting cells.

The Jaks are activated by a wide range of receptors summarized in the Table below. (Adapted from review by Schidler and Darnell, Ann. Rev. Biochem. 64:621-51 (1995)). A cytokine receptor family, capable of activating Jaks, is divided into two groups: (a) Class I includes receptors for IL-2, IL-3, IL-4, IL-6, IL-7, IL-9, IL-11, IL-12, IL-15, Epo, PRL, GH, G-CSF, GM-CSF, LIF, CNTF, and thrombopoietin; and (b) Class 2 includes IFN-a, IFN-g, and IL-10. The Class I receptors share a conserved cysteine motif (a set of four conserved cysteines and one tryptophan) and a WSXWS motif (a membrane proximal region encoding Tip-Ser-Xaa-Trp-Ser (SEQ ID NO: 2)).

Thus, on binding of a ligand to a receptor, Jaks are activated, which in turn activate STATs, which then translocate and bind to GAS elements. This entire process is encompassed in the Jaks-STATs signal transduction pathway. Therefore, activation of the Jaks-STATs pathway, reflected by the binding of the GAS or the ISRE element, can be used to indicate proteins involved in the proliferation and differentiation of cells. For example, growth factors and cytokines are known to activate the Jaks-STATs pathway (See Table below). Thus, by using GAS elements linked to reporter molecules, activators of the Jaks-STATs pathway can be identified. JAKs Ligand tyk2 Jak1 Jak2 Jak3 STATS GAS(elements) or ISRE IFN family IFN-a/B + + − − 1, 2, 3 ISRE IFN-g + + − 1 GAS (IRF1 > Lys6 > IFP) Il-10 + ? ? − 1, 3 gp130 family IL-6 (Pleiotropic) + + + ? 1, 3 GAS (IRF1 > Lys6 > IFP) Il-11 (Pleiotropic) ? + ? ? 1, 3 OnM (Pleiotropic) ? + + ? 1, 3 LIF (Pleiotropic) ? + + ? 1, 3 CNTF (Pleiotropic) −/+ + + ? 1, 3 G-CSF (Pleiotropic) ? + ? ? 1, 3 IL-12 (Pleiotropic) + − + + 1, 3 g-C family IL-2 (lymphocytes) − + − + 1, 3, 5 GAS IL-4 (lymph/myeloid) − + − + 6 GAS (IRF1 = IFP >> Ly6)(IgH) IL-7 (lymphocytes) − + − + 5 GAS IL-9 (lymphocytes) − + − + 5 GAS IL-13 (lymphocyte) − + ? ? 6 GAS IL-15 ? + ? + 5 GAS gp140 family IL-3 (myeloid) − − + − 5 GAS (IRF1 > IFP >> Ly6) IL-5 (myeloid) − − + − 5 GAS GM-CSF (myeloid) − − + − 5 GAS Growth hormone family GH ? − + − 5 PRL ? +/− + − 1, 3, 5 EPO ? − + − 5 GAS(B-CAS > IRF1 = IFP >> Ly6) Receptor Tyrosine Kinases EGF ? + + − 1, 3 GAS (IRF1) PDGF ? + + − 1, 3 CSF-1 ? + + − 1, 3 GAS (not IRF1)

To construct a synthetic GAS containing promoter element, which is used in the Biological Assays described in Examples 32-33, a PCR based strategy is employed to generate a GAS-SV40 promoter sequence. The 5′ primer contains four tandem copies of the GAS binding site found in the IRF1 promoter and previously demonstrated to bind STATs upon induction with a range of cytokines (Rothman et al., Immunity 1:457-468 (1994).), although other GAS or ISRE elements can be used instead. The 5′ primer also contains 18 bp of sequence complementary to the SV40 early promoter sequence and is flanked with an XhoI site. The sequence of the 5′ primer is: (SEQ ID NO: 3) 5′:GCGCCTCGAGATTTCCCCGAAATCTAGATTTCCCCGAAATGATTTCC CCGAAATGATTTCCCCGAAATATCTGCCATCTCAATTAG:3′

The downstream primer is complementary to the SV40 promoter and is flanked with a Hind III site: 5′:GCGGCAAGCTTTTTGCAAAGCCTAGGC:3′ (SEQ ID NO: 4)

PCR amplification is performed using the SV40 promoter template present in the B-gal:promoter plasmid obtained from Clontech. The resulting PCR fragment is digested with XhoI/Hind III and subcloned into BLSK2-. (Stratagene.) Sequencing with forward and reverse primers confirms that the insert contains the following sequence: (SEQ ID NO: 5) 5′:CTCGAGATTTCCCCGAAATCTAGATTTCCCCGAAATGATTTCCCCGA AATGATTTCCCCGAAATATCTGCCATCTCAATTAGTCAGCAACCATAGTC CCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCA TTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGG CCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGA GGCCTAGGCTTTTGCAAAAAGCTT:3′

With this GAS promoter element linked to the SV40 promoter, a GAS:SEAP2 reporter construct is next engineered. Here, the reporter molecule is a secreted alkaline phosphatase, or “SEAP.” Clearly, however, any reporter molecule can be instead of SEAP, in this or in any of the other Examples. Well known reporter molecules that can be used instead of SEAP include chloramphenicol acetyltransferase (CAT), luciferase, alkaline phosphatase, B-galactosidase, green fluorescent protein (GFP), or any protein detectable by an antibody.

The above sequence confirmed synthetic GAS-SV40 promoter element is subcloned into the pSEAP-Promoter vector obtained from Clontech using HindIII and XhoI, effectively replacing the SV40 promoter with the amplified GAS:SV40 promoter element, to create the GAS-SEAP vector. However, this vector does not contain a neomycin resistance gene, and therefore, is not preferred for mammalian expression systems.

Thus, in order to generate mammalian stable cell lines expressing the GAS-SEAP reporter, the GAS-SEAP cassette is removed from the GAS-SEAP vector using SalI and NotI, and inserted into a backbone vector containing the neomycin resistance gene, such as pGFP-1 (Clontech), using these restriction sites in the multiple cloning site, to create the GAS-SEAP/Neo vector. Once this vector is transfected into mammalian cells, this vector can then be used as a reporter molecule for GAS binding as described in Examples 32-33.

Other constructs can be made using the above description and replacing GAS with a different promoter sequence. For example, construction of reporter molecules containing EGR and NF-KB promoter sequences are described in Examples 34 and 35. However, many other promoters can be substituted using the protocols described in these Examples. For instance, SRE, IL-2, NFAT, or Osteocalcin promoters can be substituted, alone or in combination (e.g., GAS/NF-KB/EGR, GAS/NF-KB, Il-2/NFAT, or NF-KB/GAS). Similarly, other cell lines can be used to test reporter construct activity, such as HELA (epithelial), HUVEC (endothelial), Reh (B-cell), Saos-2 (osteoblast), HUVAC (aortic), or Cardiomyocyte.

Example 32 High-Throughput Screening Assay for T-Cell Activity

The following protocol is used to assess T-cell activity by identifying factors, and determining whether supernate containing a polypeptide of the invention proliferates and/or differentiates T-cells. T-cell activity is assessed using the GAS/SEAP/Neo construct produced in Example 31. Thus, factors that increase SEAP activity indicate the ability to activate the Jaks-STATS signal transduction pathway. The T-cell used in this assay is Jurkat T-cells (ATCC Accession No. TIB-152), although Molt-3 cells (ATCC Accession No. CRL-1552) and Molt-4 cells (ATCC Accession No. CRL-1582) cells can also be used.

Jurkat T-cells are lymphoblastic CD4+ Th1 helper cells. In order to generate stable cell lines, approximately 2 million Jurkat cells are transfected with the GAS-SEAP/neo vector using DMRIE-C (Life Technologies)(transfection procedure described below). The transfected cells are seeded to a density of approximately 20,000 cells per well and transfectants resistant to 1 mg/ml genticin selected. Resistant colonies are expanded and then tested for their response to increasing concentrations of interferon gamma. The dose response of a selected clone is demonstrated.

Specifically, the following protocol will yield sufficient cells for 75 wells containing 200 ul of cells. Thus, it is either scaled up, or performed in multiple to generate sufficient cells for multiple 96 well plates. Jurkat cells are maintained in RPMI+10% serum with 1% Pen-Strep. Combine 2.5 mls of OPTI-MEM (Life Technologies) with 10 ug of plasmid DNA in a T25 flask. Add 2.5 ml OPTI-MEM containing 50 ul of DMRE-C and incubate at room temperature for 15-45 mins.

During the incubation period, count cell concentration, spin down the required number of cells (10⁷ per transfection), and resuspend in OPTI-MEM to a final concentration of 10⁷ cells/ml. Then add 1 ml of 1×10⁷ cells in OPTI-MEM to T25 flask and incubate at 37 degree C. for 6 hrs. After the incubation, add 10 ml of RPMI+15% serum.

The Jurkat:GAS-SEAP stable reporter lines are maintained in RPMI+10% serum, 1 mg/ml Genticin, and 1% Pen-Strep. These cells are treated with supernatants containing polypeptide of the present invention or polypeptide of the present invention induced polypeptides as produced by the protocol described in Example 30.

On the day of treatment with the supernatant, the cells should be washed and resuspended in fresh RPMI+10% serum to a density of 500,000 cells per ml. The exact number of cells required will depend on the number of supernatants being screened. For one 96 well plate, approximately 10 million cells (for 10 plates, 100 million cells) are required.

Transfer the cells to a triangular reservoir boat, in order to dispense the cells into a 96 well dish, using a 12 channel pipette. Using a 12 channel pipette, transfer 200 ul of cells into each well (therefore adding 100,000 cells per well).

After all the plates have been seeded, 50 ul of the supernatants are transferred directly from the 96 well plate containing the supernatants into each well using a 12 channel pipette. In addition, a dose of exogenous interferon gamma (0.1, 1.0, 10 ng) is added to wells H9, H10, and H11 to serve as additional positive controls for the assay.

The 96 well dishes containing Jurkat cells treated with supernatants are placed in an incubator for 48 hrs (note: this time is variable between 48-72 hrs). 35 ul samples from each well are then transferred to an opaque 96 well plate using a 12 channel pipette. The opaque plates should be covered (using sellophene covers) and stored at −20 degree C. until SEAP assays are performed according to Example 36. The plates containing the remaining treated cells are placed at 4 degree C. and serve as a source of material for repeating the assay on a specific well if desired.

As a positive control, 100 Unit/ml interferon gamma can be used which is known to activate Jurkat T cells. Over 30 fold induction is typically observed in the positive control wells.

The above protocol may be used in the generation of both transient, as well as, stable transfected cells, which would be apparent to those of skill in the art.

Example 33 High-Throughput Screening Assay Identifying Myeloid Activity

The following protocol is used to assess myeloid activity of polypeptide of the present invention by determining whether polypeptide of the present invention proliferates and/or differentiates myeloid cells. Myeloid cell activity is assessed using the GAS/SEAP/Neo construct produced in Example 31. Thus, factors that increase SEAP activity indicate the ability to activate the Jaks-STATS signal transduction pathway. The myeloid cell used in this assay is U937, a pre-monocyte cell line, although TF-1, HL60, or KG1 can be used.

To transiently transfect U937 cells with the GAS/SEAP/Neo construct produced in Example 31, a DEAE-Dextran method (Kharbanda et. al., 1994, Cell Growth & Differentiation, 5:259-265) is used. First, harvest 2×10⁷ U937 cells and wash with PBS. The U937 cells are usually grown in RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum (FBS) supplemented with 100 units/ml penicillin and 100 mg/ml streptomycin.

Next, suspend the cells in 1 ml of 20 mM Tris-HCl (pH 7.4) buffer containing 0.5 mg/ml DEAE-Dextran, 8 ug GAS-SEAP2 plasmid DNA, 140 mM NaCl, 5 mM KCl, 375 uM Na₂BPO₄.7H₂O, 1 mM MgCl₂, and 675 uM CaCl₂. Incubate at 37 degrees C. for 45 min.

Wash the cells with RPMI 1640 medium containing 10% FBS and then resuspend in 10 ml complete medium and incubate at 37 degree C. for 36 hr.

The GAS-SEAP/U937 stable cells are obtained by growing the cells in 400 ug/ml G418. The G418-free medium is used for routine growth but every one to two months, the cells should be re-grown in 400 ug/ml G418 for couple of passages.

These cells are tested by harvesting 1×10⁸ cells (this is enough for ten 96-well plates assay) and wash with PBS. Suspend the cells in 200 ml above described growth medium, with a final density of 5×10⁵ cells/ml. Plate 200 ul cells per well in the 96-well plate (or 1×10⁵ cells/well).

Add 50 ul of the supernatant prepared by the protocol described in Example 30. Incubate at 37 degree C. for 48 to 72 hr. As a positive control, 100 Unit/ml interferon gamma can be used which is known to activate U937 cells. Over 30 fold induction is typically observed in the positive control wells. SEAP assay the supernatant according to the protocol described in Example 36.

Example 34 High-Throughput Screening Assay Identifying Neuronal Activity

When cells undergo differentiation and proliferation, a group of genes are activated through many different signal transduction pathways. One of these genes, EGR1 (early growth response gene 1), is induced in various tissues and cell types upon activation. The promoter of EGR1 is responsible for such induction. Using the EGR1 promoter linked to reporter molecules, activation of cells can be assessed by polypeptide of the present invention.

Particularly, the following protocol is used to assess neuronal activity in PC12 cell lines. PC12 cells (rat phenochromocytoma cells) are known to proliferate and/or differentiate by activation with a number of mitogens, such as TPA (tetradecanoyl phorbol acetate), NGF (nerve growth factor), and EGF (epidermal growth factor). The EGR1 gene expression is activated during this treatment. Thus, by stably transfecting PC12 cells with a construct containing an EGR promoter linked to SEAP reporter, activation of PC12 cells by polypeptide of the present invention can be assessed.

The EGR/SEAP reporter construct can be assembled by the following protocol. The EGR-1 promoter sequence (−633 to +1)(Sakamoto K et al., Oncogene 6:867-871 (1991)) can be PCR amplified from human genomic DNA using the following primers: (SEQ ID NO: 6) 5′ GCGCTCGAGGGATGACAGCGATAGAACCCCGG-3′ (SEQ ID NO: 7) 5′ GCGAAGCTTCGCGACTCCCCGGATCCGCCTC-3′

Using the GAS:SEAP/Neo vector produced in Example 31, EGR1 amplified product can then be inserted into this vector. Linearize the GAS:SEAP/Neo vector using restriction enzymes XhoI/HindIII, removing the GAS/SV40 stuffer. Restrict the EGR1 amplified product with these same enzymes. Ligate the vector and the EGR1 promoter.

To prepare 96 well-plates for cell culture, two mls of a coating solution (1:30 dilution of collagen type I (Upstate Biotech Inc. Cat#08-115) in 30% ethanol (filter sterilized)) is added per one 10 cm plate or 50 ml per well of the 96-well plate, and allowed to air dry for 2 hr.

PC12 cells are routinely grown in RPMI-1640 medium (Bio Whittaker) containing 10% horse serum (JRH BIOSCIENCES, Cat. # 12449-78P), 5% heat-inactivated fetal bovine serum (FBS) supplemented with 100 units/ml penicillin and 100 ug/ml streptomycin on a precoated 10 cm tissue culture dish. One to four split is done every three to four days. Cells are removed from the plates by scraping and resuspended with pipetting up and down for more than 15 times.

Transfect the EGR/SEAP/Neo construct into PC12 using the Lipofectamine protocol described in Example 30. EGR-SEAP/PC12 stable cells are obtained by growing the cells in 300 ug/ml G418. The G418-free medium is used for routine growth but every one to two months, the cells should be re-grown in 300 ug/ml G418 for couple of passages.

To assay for neuronal activity, a 10 cm plate with cells around 70 to 80% confluent is screened by removing the old medium. Wash the cells once with PBS (Phosphate buffered saline). Then starve the cells in low serum medium (RPMI-1640 containing 1% horse serum and 0.5% FBS with antibiotics) overnight.

The next morning, remove the medium and wash the cells with PBS. Scrape off the cells from the plate, suspend the cells well in 2 ml low serum medium. Count the cell number and add more low serum medium to reach final cell density as 5×10⁵ cells/ml.

Add 200 ul of the cell suspension to each well of 96-well plate (equivalent to 1×10⁵ cells/well). Add 50 ul supernatant produced by Example 30, 37 degree C. for 48 to 72 hr. As a positive control, a growth factor known to activate PC12 cells through EGR can be used, such as 50 ng/ul of Neuronal Growth Factor (NGF). Over fifty-fold induction of SEAP is typically seen in the positive control wells. SEAP assay the supernatant according to Example 36.

Example 35 High-Throughput Screening Assay for T-Cell Activity

NF-KB (Nuclear Factor KB) is a transcription factor activated by a wide variety of agents including the inflammatory cytokines IL-1 and TNF, CD30 and CD40, lymphotoxin-alpha and lymphotoxin-beta, by exposure to LPS or thrombin, and by expression of certain viral gene products. As a transcription factor, NF-KB regulates the expression of genes involved in immune cell activation, control of apoptosis (NF-KB appears to shield cells from apoptosis), B and T-cell development, anti-viral and antimicrobial responses, and multiple stress responses.

In non-stimulated conditions, NF-KB is retained in the cytoplasm with I-KB (Inhibitor KB). However, upon stimulation, I-KB is phosphorylated and degraded, causing NF-KB to shuttle to the nucleus, thereby activating transcription of target genes. Target genes activated by NF-KB include IL-2, IL-6, GM-CSF, ICAM-1 and class I MHC.

Due to its central role and ability to respond to a range of stimuli, reporter constructs utilizing the NF-KB promoter element are used to screen the supernatants produced in Example 30. Activators or inhibitors of NF-KB would be useful in treating, preventing, and/or diagnosing diseases. For example, inhibitors of NF-KB could be used to treat those diseases related to the acute or chronic activation of NF-KB, such as rheumatoid arthritis.

To construct a vector containing the NF-KB promoter element, a PCR based strategy is employed. The upstream primer contains four tandem copies of the NF-KB binding site (GGGGACTTTCCC) (SEQ ID NO: 8), 18 bp of sequence complementary to the 5′ end of the SV40 early promoter sequence, and is flanked with an XhoI site: (SEQ ID NO: 9) 5′:GCGGCCTCGAGGGGACTTTCCCGGGGACTTTCCGGGGACTTTCCGGG ACTTTCCATCCTGCCATCTCAATTAG:3′

The downstream primer is complementary to the 3′ end of the SV40 promoter and is flanked with a Hind III site: (SEQ ID NO: 4) 5′:GCGGCAAGCTTTTTGCAAAGCCTAGGC:3′

PCR amplification is performed using the SV40 promoter template present in the pB-gal:promoter plasmid obtained from Clontech. The resulting PCR fragment is digested with XhoI and Hind III and subcloned into BLSK2-. (Stratagene) Sequencing with the T7 and T3 primers confirms the insert contains the following sequence: (SEQ ID NO: 10) 5′:CTCGAGGGGACTTTCCCGGGGACTTTCCGGGGACTTTCCGGGACTTT CCATCTGCCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCG CCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGG CTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTG AGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGC AAAAAGCTT:3′

Next, replace the SV40 minimal promoter element present in the pSEAP2-promoter plasmid (Clontech) with this NF-KB/SV40 fragment using XhoI and HindIII. However, this vector does not contain a neomycin resistance gene, and therefore, is not preferred for mammalian expression systems.

In order to generate stable mammalian cell lines, the NF-KB/SV40/SEAP cassette is removed from the above NF-KB/SEAP vector using restriction enzymes SalI and NotI, and inserted into a vector containing neomycin resistance. Particularly, the NF-KB/SV40/SEAP cassette was inserted into pGFP-1 (Clontech), replacing the GFP gene, after restricting pGFP-1 with SalI and NotI.

Once NF-KB/SV40/SEAP/Neo vector is created, stable Jurkat T-cells are created and maintained according to the protocol described in Example 32. Similarly, the method for assaying supernatants with these stable Jurkat T-cells is also described in Example 32. As a positive control, exogenous TNF alpha (0.1, 1, 10 ng) is added to wells H9, H10, and H11, with a 5-10 fold activation typically observed.

Example 36 Assay for SEAP Activity

As a reporter molecule for the assays described in Examples 32-35, SEAP activity is assayed using the Tropix Phospho-light Kit (Cat. BP-400) according to the following general procedure. The Tropix Phospho-light Kit supplies the Dilution, Assay, and Reaction Buffers used below.

Prime a dispenser with the 2.5× Dilution Buffer and dispense 15 ul of 2.5× dilution buffer into Optiplates containing 35 ul of a supernatant. Seal the plates with a plastic sealer and incubate at 65 degree C. for 30 min. Separate the Optiplates to avoid uneven heating.

Cool the samples to room temperature for 15 minutes. Empty the dispenser and prime with the Assay Buffer. Add 50 ml Assay Buffer and incubate at room temperature 5 min. Empty the dispenser and prime with the Reaction Buffer (see the Table below). Add 50 ul Reaction Buffer and incubate at room temperature for 20 minutes. Since the intensity of the chemiluminescent signal is time dependent, and it takes about 10 minutes to read 5 plates on a luminometer, thus one should treat 5 plates at each time and start the second set 10 minutes later.

Read the relative light unit in the luminometer. Set H12 as blank, and print the results. An increase in chemiluminescence indicates reporter activity. Reaction Buffer Formulation: # of plates Rxn buffer diluent (ml) CSPD (ml) 10 60 3 11 65 3.25 12 70 3.5 13 75 3.75 14 80 4 15 85 4.25 16 90 4.5 17 95 4.75 18 100 5 19 105 5.25 20 110 5.5 21 115 5.75 22 120 6 23 125 6.25 24 130 6.5 25 135 6.75 26 140 7 27 145 7.25 28 150 7.5 29 155 7.75 30 160 8 31 165 8.25 32 170 8.5 33 175 8.75 34 180 9 35 185 9.25 36 190 9.5 37 195 9.75 38 200 10 39 205 10.25 40 210 10.5 41 215 10.75 42 220 11 43 225 11.25 44 230 11.5 45 235 11.75 46 240 12 47 245 12.25 48 250 12.5 49 255 12.75 50 260 13

Example 37 High-Throughput Screening Assay Identifying Changes in Small Molecule Concentration and Membrane Permeability

Binding of a ligand to a receptor is known to alter intracellular levels of small molecules, such as calcium, potassium, sodium, and pH, as well as alter membrane potential. These alterations can be measured in an assay to identify supernatants which bind to receptors of a particular cell. Although the following protocol describes an assay for calcium, this protocol can easily be modified to detect changes in potassium, sodium, pH, membrane potential, or any other small molecule which is detectable by a fluorescent probe.

The following assay uses Fluorometric Imaging Plate Reader (“FLIPR”) to measure changes in fluorescent molecules (Molecular Probes) that bind small molecules. Clearly, any fluorescent molecule detecting a small molecule can be used instead of the calcium fluorescent molecule, fluo-4 (Molecular Probes, Inc.; catalog no. F-14202), used here.

For adherent cells, seed the cells at 10,000 cells/well in a Co-star black 96-well plate with clear bottom. The plate is incubated in a CO₂ incubator for 20 hours. The adherent cells are washed two times in Biotek washer with 200 ul of HBSS (Hank's Balanced salt Solution) leaving 100 ul of buffer after the final wash.

A stock solution of 1 mg/ml fluo-4 is made in 10% pluronic acid DMSO. To load the cells with fluo-4, 50 ul of 12 ug/ml fluo-4 is added to each well. The plate is incubated at 37 degrees C. in a CO₂ incubator for 60 min. The plate is washed four times in the Bioteck washer with HBSS leaving 100 ul of buffer.

For non-adherent cells, the cells are spun down from culture media. Cells are re-suspended to 2-5×10⁶ cells/ml with HBSS in a 50-ml conical tube. 4 ul of 1 mg/ml fluo-4 solution in 10% pluronic acid DMSO is added to each ml of cell suspension. The tube is then placed in a 37 degrees C. water bath for 30-60 min. The cells are washed twice with HBSS, resuspended to 1×10⁶ cells/ml, and dispensed into a microplate, 100 ul/well. The plate is centrifuged at 1000 rpm for 5 min. The plate is then washed once in Denley Cell Wash with 200 ul, followed by an aspiration step to 100 ul final volume.

For a non-cell based assay, each well contains a fluorescent molecule, such as fluo-4. The supernatant is added to the well, and a change in fluorescence is detected.

To measure the fluorescence of intracellular calcium, the FLIPR is set for the following parameters: (1) System gain is 300-800 mW; (2) Exposure time is 0.4 second; (3) Camera F/stop is F/2; (4) Excitation is 488 nm; (5) Emission is 530 nm; and (6) Sample addition is 50 ul. Increased emission at 530 nm indicates an extracellular signaling event caused by the a molecule, either polypeptide of the present invention or a molecule induced by polypeptide of the present invention, which has resulted in an increase in the intracellular Ca++ concentration.

Example 38 High-Throughput Screening Assay Identifying Tyrosine Kinase Activity

The Protein Tyrosine Kinases (PTK) represent a diverse group of transmembrane and cytoplasmic kinases. Within the Receptor Protein Tyrosine Kinase RPTK) group are receptors for a range of mitogenic and metabolic growth factors including the PDGF, FGF, EGF, NGF, HGF and Insulin receptor subfamilies. In addition there are a large family of RPTKs for which the corresponding ligand is unknown. Ligands for RPTKs include mainly secreted small proteins, but also membrane-bound and extracellular matrix proteins.

Activation of RPTK by ligands involves ligand-mediated receptor dimerization, resulting in transphosphorylation of the receptor subunits and activation of the cytoplasmic tyrosine kinases. The cytoplasmic tyrosine kinases include receptor associated tyrosine kinases of the src-family (e.g., src, yes, lck, lyn, fyn) and non-receptor linked and cytosolic protein tyrosine kinases, such as the Jak family, members of which mediate signal transduction triggered by the cytokine superfamily of receptors (e.g., the Interleukins, Interferons, GM-CSF, and Leptin).

Because of the wide range of known factors capable of stimulating tyrosine kinase activity, identifying whether polypeptide of the present invention or a molecule induced by polypeptide of the present invention is capable of activating tyrosine kinase signal transduction pathways is of interest. Therefore, the following protocol is designed to identify such molecules capable of activating the tyrosine kinase signal transduction pathways.

Seed target cells (e.g., primary keratinocytes) at a density of approximately 25,000 cells per well in a 96 well Loprodyne Silent Screen Plates purchased from Nalge Nunc (Naperville, Ill.). The plates are sterilized with two 30 minute rinses with 100% ethanol, rinsed with water and dried overnight. Some plates are coated for 2 hr with 100 ml of cell culture grade type I collagen (50 mg/ml), gelatin (2%) or polylysine (50 mg/ml), all of which can be purchased from Sigma Chemicals (St. Louis, Mo.) or 10% Matrigel purchased from Becton Dickinson (Bedford, Mass.), or calf serum, rinsed with PBS and stored at 4 degree C. Cell growth on these plates is assayed by seeding 5,000 cells/well in growth medium and indirect quantitation of cell number through use of alamarBlue as described by the manufacturer Alamar Biosciences, Inc. (Sacramento, Calif.) after 48 hr. Falcon plate covers #3071 from Becton Dickinson (Bedford, Mass.) are used to cover the Loprodyne Silent Screen Plates. Falcon Microtest III cell culture plates can also be used in some proliferation experiments.

To prepare extracts, A431 cells are seeded onto the nylon membranes of Loprodyne plates (20,000/200 ml/well) and cultured overnight in complete medium. Cells are quiesced by incubation in serum-free basal medium for 24 hr. After 5-20 minutes treatment with EGF (60 ng/ml) or 50 ul of the supernatant produced in Example 30, the medium was removed and 100 ml of extraction buffer ((20 mM HEPES pH 7.5, 0.15 M NaCl, 1% Triton X-100, 0.1% SDS, 2 mM Na3VO4, 2 mM Na4P2O7 and a cocktail of protease inhibitors (# 1836170) obtained from Boeheringer Mannheim (Indianapolis, Ind.)) is added to each well and the plate is shaken on a rotating shaker for 5 minutes at 4° C. The plate is then placed in a vacuum transfer manifold and the extract filtered through the 0.45 mm membrane bottoms of each well using house vacuum. Extracts are collected in a 96-well catch/assay plate in the bottom of the vacuum manifold and immediately placed on ice. To obtain extracts clarified by centrifugation, the content of each well, after detergent solubilization for 5 minutes, is removed and centrifuged for 15 minutes at 4 degree C. at 16,000×g.

Test the filtered extracts for levels of tyrosine kinase activity. Although many methods of detecting tyrosine kinase activity are known, one method is described here.

Generally, the tyrosine kinase activity of a supernatant is evaluated by determining its ability to phosphorylate a tyrosine residue on a specific substrate (a biotinylated peptide). Biotinylated peptides that can be used for this purpose include PSK1 (corresponding to amino acids 6-20 of the cell division kinase cdc2-p34) and PSK2 (corresponding to amino acids 1-17 of gastrin). Both peptides are substrates for a range of tyrosine kinases and are available from Boehringer Mannheim.

The tyrosine kinase reaction is set up by adding the following components in order. First, add 10 ul of 5 uM Biotinylated Peptide, then 10 ul ATP/Mg₂₊ (5 mM ATP/50 mM MgCl₂), then 10 ul of 5× Assay Buffer (40 mM imidazole hydrochloride, pH7.3, 40 mM beta-glycerophosphate, 1 mM EGTA, 100 mM MgCl₂, 5 mM MnCl₂, 0.5 mg/ml BSA), then 5 ul of Sodium Vanadate (1 mM), and then 5 ul of water. Mix the components gently and preincubate the reaction mix at 30 degree C. for 2 min. Initial the reaction by adding 10 ul of the control enzyme or the filtered supernatant.

The tyrosine kinase assay reaction is then terminated by adding 10 ul of 120 mm EDTA and place the reactions on ice.

Tyrosine kinase activity is determined by transferring 50 ul aliquot of reaction mixture to a microtiter plate (MTP) module and incubating at 37 degree C. for 20 min. This allows the streptavidin coated 96 well plate to associate with the biotinylated peptide. Wash the MTP module with 300 ul/well of PBS four times. Next add 75 ul of anti-phospotyrosine antibody conjugated to horse radish peroxidase (anti-P-Tyr-POD (0.5 u/ml)) to each well and incubate at 37 degree C. for one hour. Wash the well as above.

Next add 100 ul of peroxidase substrate solution (Boehringer Mannheim) and incubate at room temperature for at least 5 mins (up to 30 min). Measure the absorbance of the sample at 405 nm by using ELISA reader. The level of bound peroxidase activity is quantitated using an ELISA reader and reflects the level of tyrosine kinase activity.

Example 39 High-Throughput Screening Assay Identifying Phosphorylation Activity

As a potential alternative and/or complement to the assay of protein tyrosine kinase activity described in Example 38, an assay which detects activation (phosphorylation) of major intracellular signal transduction intermediates can also be used. For example, as described below one particular assay can detect tyrosine phosphorylation of the Erk-1 and Erk-2 kinases. However, phosphorylation of other molecules, such as Raf, JNK, p38 MAP, Map kinase kinase (MEK), MEK kinase, Src, Muscle specific kinase (MuSK), IRAK, Tec, and Janus, as well as any other phosphoserine, phosphotyrosine, or phosphothreonine molecule, can be detected by substituting these molecules for Erk-1 or Erk-2 in the following assay.

Specifically, assay plates are made by coating the wells of a 96-well ELISA plate with 0.1 ml of protein G (1 ug/ml) for 2 hr at room temp, (RT). The plates are then rinsed with PBS and blocked with 3% BSA/PBS for 1 hr at RT. The protein G plates are then treated with 2 commercial monoclonal antibodies (100 ng/well) against Erk-1 and Erk-2 (1 hr at RT) (Santa Cruz Biotechnology). (To detect other molecules, this step can easily be modified by substituting a monoclonal antibody detecting any of the above described molecules.) After 3-5 rinses with PBS, the plates are stored at 4 degree C. until use.

A431 cells are seeded at 20,000/well in a 96-well Loprodyne filterplate and cultured overnight in growth medium. The cells are then starved for 48 hr in basal medium (DMEM) and then treated with EGF (6 ng/well) or 50 ul of the supernatants obtained in Example 30 for 5-20 minutes. The cells are then solubilized and extracts filtered directly into the assay plate.

After incubation with the extract for 1 hr at RT, the wells are again rinsed. As a positive control, a commercial preparation of MAP kinase (10 ng/well) is used in place of A431 extract. Plates are then treated with a commercial polyclonal (rabbit) antibody (1 ug/ml) which specifically recognizes the phosphorylated epitope of the Erk-1 and Erk-2 kinases (1 hr at RT). This antibody is biotinylated by standard procedures. The bound polyclonal antibody is then quantitated by successive incubations with Europium-streptavidin and Europium fluorescence enhancing reagent in the Wallac DELFIA instrument (time-resolved fluorescence). An increased fluorescent signal over background indicates a phosphorylation by polypeptide of the present invention or a molecule induced by polypeptide of the present invention.

Example 40 Assay for the Stimulation of Bone Marrow CD34+ Cell Proliferation

This assay is based on the ability of human CD34+ to proliferate in the presence of hematopoietic growth factors and evaluates the ability of isolated polypeptides expressed in mammalian cells to stimulate proliferation of CD34+ cells.

It has been previously shown that most mature precursors will respond to only a single signal. More immature precursors require at least two signals to respond. Therefore, to test the effect of polypeptides on hematopoietic activity of a wide range of progenitor cells, the assay contains a given polypeptide in the presence or absence of other hematopoietic growth factors. Isolated cells are cultured for 5 days in the presence of Stem Cell Factor (SCF) in combination with tested sample. SCF alone has a very limited effect on the proliferation of bone marrow (BM) cells, acting in such conditions only as a “survival” factor. However, combined with any factor exhibiting stimulatory effect on these cells (e.g., IL-3), SCF will cause a synergistic effect. Therefore, if the tested polypeptide has a stimulatory effect on hematopoietic progenitors, such activity can be easily detected. Since normal BM cells have a low level of cycling cells, it is likely that any inhibitory effect of a given polypeptide, or agonists or antagonists thereof, might not be detected. Accordingly, assays for an inhibitory effect on progenitors is preferably tested in cells that are first subjected to in vitro stimulation with SCF+IL+3, and then contacted with the compound that is being evaluated for inhibition of such induced proliferation.

Briefly, CD34+ cells are isolated using methods known in the art. The cells are thawed and resuspended in medium (QBSF 60 serum-free medium with 1% L-glutamine (500 ml) Quality Biological, Inc., Gaithersburg, Md. Cat# 160-204-101). After several gentle centrifugation steps at 200×g, cells are allowed to rest for one hour. The cell count is adjusted to 2.5×10⁵ cells/ml. During this time, 100 μl of sterile water is added to the peripheral wells of a 96-well plate. The cytokines that can be tested with a given polypeptide in this assay is rhSCF (R&D Systems, Minneapolis, Minn., Cat# 255-SC) at 50 ng/ml alone and in combination with rhSCF and rhIL-3 (R&D Systems, Minneapolis, Minn., Cat# 203-ML) at 30 ng/ml. After one hour, 10 μl of prepared cytokines, 50 μl of the supernatants prepared in Example 30 (supernatants at 1:2 dilution=50 μl) and 20 μl of diluted cells are added to the media which is already present in the wells to allow for a final total volume of 100 μl. The plates are then placed in a 37° C./5% CO₂ incubator for five days.

Eighteen hours before the assay is harvested, 0.5 μCi/well of [3H] Thymidine is added in a 10 μl volume to each well to determine the proliferation rate. The experiment is terminated by harvesting the cells from each 96-well plate to a filtermat using the Tomtec Harvester 96. After harvesting, the filtermats are dried, trimmed and placed into OmniFilter assemblies consisting of one OmniFilter plate and one OmniFilter Tray. 60 μl Microscint is added to each well and the plate sealed with TopSeal-A press-on sealing film A bar code 15 sticker is affixed to the first plate for counting. The sealed plates are then loaded and the level of radioactivity determined via the Packard Top Count and the printed data collected for analysis. The level of radioactivity reflects the amount of cell proliferation.

The studies described in this example test the activity of a given polypeptide to stimulate bone marrow CD34+ cell proliferation. One skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof. As a nonlimiting example, potential antagonists tested in this assay would be expected to inhibit cell proliferation in the presence of cytokines and/or to increase the inhibition of cell proliferation in the presence of cytokines and a given polypeptide. In contrast, potential agonists tested in this assay would be expected to enhance cell proliferation and/or to decrease the inhibition of cell proliferation in the presence of cytokines and a given polypeptide.

The ability of a gene to stimulate the proliferation of bone marrow CD34+ cells indicates that polynucleotides and polypeptides corresponding to the gene are useful for the diagnosis and treatment of disorders affecting the immune system and hematopoiesis. Representative uses are described in the “Immune Activity” and “Infectious Disease” sections above, and elsewhere herein.

Example 41 Assay for Extracellular Matrix Enhanced Cell Response (EMECR)

The objective of the Extracellular Matrix Enhanced Cell Response (EMECR) assay is to identify gene products (e.g., isolated polypeptides) that act on the hematopoietic stem cells in the context of the extracellular matrix (ECM) induced signal.

Cells respond to the regulatory factors in the context of signal(s) received from the surrounding microenvironment. For example, fibroblasts, and endothelial and epithelial stem cells fail to replicate in the absence of signals from the ECM. Hematopoietic stem cells can undergo self-renewal in the bone marrow, but not in in vitro suspension culture. The ability of stem cells to undergo self-renewal in vitro is dependent upon their interaction with the stromal cells and the ECM protein fibronectin (fn). Adhesion of cells to fn is mediated by the α₅.β₁ and α₄.β₁ integrin receptors, which are expressed by human and mouse hematopoietic stem cells. The factor(s) which integrate with the ECM environment and are responsible for stimulating stem cell self-renewal have a not yet been identified. Discovery of such factors should be of great interest in gene therapy and bone marrow transplant applications

Briefly, polystyrene, non tissue culture treated, 96-well plates are coated with fn fragment at a coating concentration of 0.2 μg/cm². Mouse bone marrow cells are plated (1,000 cells/well) in 0.2 ml of serum-free medium. Cells cultured in the presence of IL-3 (5 ng/ml)+SCF (50 ng/ml) would serve as the positive control, conditions under which little self-renewal but pronounced differentiation of the stem cells is to be expected. Gene products of the invention (e.g., including, but not limited to, polynucleotides and polypeptides of the present invention, and supernatants produced in Example 30), are tested with appropriate negative controls in the presence and absence of SCF (5.0 ng/ml), where test factor supernatants represent 10% of the total assay volume. The plated cells are then allowed to grow by incubating in a low oxygen environment (5% CO₂, 7% O₂, and 88% N₂) tissue culture incubator for 7 days. The number of proliferating cells within the wells is then quantitated by measuring thymidine incorporation into cellular DNA. Verification of the positive hits in the assay will require phenotypic characterization of the cells, which can be accomplished by scaling up of the culture system and using appropriate antibody reagents against cell surface antigens and FACScan.

One skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof.

If a particular polypeptide of the present invention is found to be a stimulator of hematopoietic progenitors, polynucleotides and polypeptides corresponding to the gene encoding said polypeptide may be useful for the diagnosis and treatment of disorders affecting the immune system and hematopoiesis. Representative uses are described in the “Immune Activity” and “Infectious Disease” sections above, and elsewhere herein. The gene product may also be useful in the expansion of stem cells and committed progenitors of various blood lineages, and in the differentiation and/or proliferation of various cell types.

Additionally, the polynucleotides and/or polypeptides of the gene of interest and/or agonists and/or antagonists thereof, may also be employed to inhibit the proliferation and differentiation of hematopoietic cells and therefore may be employed to protect bone marrow stem cells from chemotherapeutic agents during chemotherapy. This antiproliferative effect may allow administration of higher doses of chemotherapeutic agents and, therefore, more effective chemotherapeutic treatment.

Moreover, polynucleotides and polypeptides corresponding to the gene of interest may also be useful for the treatment and diagnosis of hematopoietic related disorders such as, for example, anemia, pancytopenia, leukopenia, thrombocytopenia or leukemia since stromal cells are important in the production of cells of hematopoietic lineages. The uses include bone marrow cell ex-vivo culture, bone marrow transplantation, bone marrow reconstitution, radiotherapy or chemotherapy of neoplasia.

Example 42 Human Dermal Fibroblast and Aortic Smooth Muscle Cell Proliferation

The polypeptide of interest is added to cultures of normal human dermal fibroblasts (NHDF) and human aortic smooth muscle cells (AoSMC) and two co-assays are performed with each sample. The first assay examines the effect of the polypeptide of interest on the proliferation of normal human dermal fibroblasts (NHDF) or aortic smooth muscle cells (AoSMC). Aberrant growth of fibroblasts or smooth muscle cells is a part of several pathological processes, including fibrosis, and restenosis. The second assay examines IL6 production by both NHDF and SMC. IL6 production is an indication of functional activation. Activated cells will have increased production of a number of cytokines and other factors, which can result in a proinflammatory or immunomodulatory outcome. Assays are run with and without co-TNFa stimulation, in order to check for costimulatory or inhibitory activity.

Briefly, on day 1,96-well black plates are set up with 1000 cells/well (NHDF) or 2000 cells/well (AOSMC) in 100 μl culture media. NHDF culture media contains: Clonetics FB basal media, 1 mg/ml hFGF, 5 mg/ml insulin, 50 mg/ml gentamycin, 2% FBS, while AoSMC culture media contains Clonetics SM basal media, 0.5 μg/ml hEGF, 5 mg/ml insulin, 1 μg/ml hFGF, 50 mg/ml gentamycin, 50 μg/ml Amphotericin B, 5% FBS. After incubation at 37° C. for at least 4-5 hours culture media is aspirated and replaced with growth arrest media. Growth arrest media for NHDF contains fibroblast basal media, 50 mg/ml gentamycin, 2% FBS, while growth arrest media for AoSMC contains SM basal media, 50 mg/ml gentamycin, 50 μg/ml Amphotericin B, 0.4% FBS. Incubate at 37° C. until day 2.

On day 2, serial dilutions and templates of the polypeptide of interest are designed such that they always include media controls and known-protein controls. For both stimulation and inhibition experiments, proteins are diluted in growth arrest media. For inhibition experiments, TNFa is added to a final concentration of 2 ng/ml (NHDF) or 5 ng/ml (AoSMC). Add ⅓ vol media containing controls or polypeptides of the present invention and incubate at 37 degrees C./5% CO₂ until day 5.

Transfer 60 μl from each well to another labeled 96-well plate, cover with a plate-sealer, and store at 4 degrees C. until Day 6 (for IL6 ELISA). To the remaining 100 μl in the cell culture plate, aseptically add Alamar Blue in an amount equal to 10% of the culture volume (10 μl). Return plates to incubator for 3 to 4 hours. Then measure fluorescence with excitation at 530 nm and emission at 590 nm using the CytoFluor. This yields the growth stimulation/inhibition data.

On day 5, the IL6 ELISA is performed by coating a 96 well plate with 50-100 ul/well of Anti-Human IL6 Monoclonal antibody diluted in PBS, pH 7.4, incubate ON at room temperature.

On day 6, empty the plates into the sink and blot on paper towels. Prepare Assay Buffer containing PBS with 4% BSA. Block the plates with 2001 μl/well of Pierce Super Block blocking buffer in PBS for 1-2 hr and then wash plates with wash buffer (PBS, 0.05% Tween-20). Blot plates on paper towels. Then add 50 μl/well of diluted Anti-Human IL-6 Monoclonal, Biotin-labeled antibody at 0.50 mg/ml. Make dilutions of IL-6 stock in media (30, 10, 3, 1, 0.3, 0 ng/ml). Add duplicate samples to top row of plate. Cover the plates and incubate for 2 hours at RT on shaker.

Plates are washed with wash buffer and blotted on paper towels. Dilute EU-labeled Streptavidin 1:1000 in Assay buffer, and add 100 μl/well. Cover the plate and incubate 1 h at RT. Plates are again washed with wash buffer and blotted on paper towels.

Add 100 μl/well of Enhancement Solution. Shake for 5 minutes. Read the plate on the Wallac DELFIA Fluorometer. Readings from triplicate samples in each assay were tabulated and averaged.

A positive result in this assay suggests AoSMC cell proliferation and that the polypeptide of the present invention may be involved in dermal fibroblast proliferation and/or smooth muscle cell proliferation. A positive result also suggests many potential uses of polypeptides, polynucleotides, agonists and/or antagonists of the polynucleotide/polypeptide of the present invention which gives a positive result. For example, inflammation and immune responses, wound healing, and angiogenesis, as detailed throughout this specification. Particularly, polypeptides of the present invention and polynucleotides of the present invention may be used in wound healing and dermal regeneration, as well as the promotion of vasculogenesis, both of the blood vessels and lymphatics. The growth of vessels can be used in the treatment of, for example, cardiovascular diseases. Additionally, antagonists of polypeptides and polynucleotides of the invention may be useful in treating diseases, disorders, and/or conditions which involve angiogenesis by acting as an anti-vascular agent (e.g., anti-angiogenesis). These diseases, disorders, and/or conditions are known in the art and/or are described herein, such as, for example, malignancies, solid tumors, benign tumors, for example hemangiomas, acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas; artheroscleric plaques; ocular angiogenic diseases, for example, diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis, retinoblastoma, uvietis and Pterygia (abnormal blood vessel growth) of the eye; rheumatoid arthritis; psoriasis; delayed wound healing; endometriosis; vasculogenesis; granulations; hypertrophic scars (keloids); nonunion fractures; scleroderma; trachoma; vascular adhesions; myocardial angiogenesis; coronary collaterals; cerebral collaterals; arteriovenous malformations; ischemic limb angiogenesis; Osler-Webber Syndrome; plaque neovascularization; telangiectasia; hemophiliac joints; angiofibroma; fibromuscular dysplasia; wound granulation; Crohn's disease; and atherosclerosis. Moreover, antagonists of polypeptides and polynucleotides of the invention may be useful in treating anti-hyperproliferative diseases and/or anti-inflammatory known in the art and/or described herein.

One skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof.

Example 43 Cellular Adhesion Molecule (CAM) Expression on Endothelial Cells

The recruitment of lymphocytes to areas of inflammation and angiogenesis involves specific receptor-ligand interactions between cell surface adhesion molecules (CAMs) on lymphocytes and the vascular endothelium. The adhesion process, in both normal and pathological settings, follows a multi-step cascade that involves intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial leukocyte adhesion molecule-1 (E-selectin) expression on endothelial cells (EC). The expression of these molecules and others on the vascular endothelium determines the efficiency with which leukocytes may adhere to the local vasculature and extravasate into the local tissue during the development of an inflammatory response. The local concentration of cytokines and growth factor participate in the modulation of the expression of these CAMs.

Briefly, endothelial cells (e.g., Human Umbilical Vein Endothelial cells (HUVECs)) are grown in a standard 96 well plate to confluence, growth medium is removed from the cells and replaced with 100 μl of 199 Medium (10% fetal bovine serum (FBS)). Samples for testing and positive or negative controls are added to the plate in triplicate (in 10 μl volumes). Plates are then incubated at 37° C. for either 5 h (selectin and integrin expression) or 24 h (integrin expression only). Plates are aspirated to remove medium and 100 μl of 0.1% paraformaldehyde-PBS (with Ca++ and Mg++) is added to each well. Plates are held at 4° C. for 30 min. Fixative is removed from the wells and wells are washed 1× with PBS(+Ca,Mg)+0.5% BSA and drained. 10 μl of diluted primary antibody is added to the test and control wells. Anti-ICAM-1-Biotin, Anti-VCAM-1-Biotin and Anti-E-selectin-Biotin are used at a concentration of 10 μg/ml (1:10 dilution of 0.1 mg/ml stock antibody). Cells are incubated at 37° C. for 30 min. in a humidified environment. Wells are washed three times with PBS(+Ca,Mg)+0.5% BSA. 20 μl of diluted ExtrAvidin-Alkaline Phosphatase (1:5,000 dilution, referred to herein as the working dilution) are added to each well and incubated at 37° C. for 30 min. Wells are washed three times with PBS(+Ca,Mg)+0.5% BSA. Dissolve 1 tablet of p-Nitrophenol Phosphate pNPP per 5 ml of glycine buffer (pH 10.4). 100 μl of pNPP substrate in glycine buffer is added to each test well. Standard wells in triplicate are prepared from the working dilution of the ExtrAvidin-Alkaline Phosphotase in glycine buffer: 1:5,000 (10⁰)>10^(−0.5)>10^(−1>10) ^(−1.5)0.5 μl of each dilution is added to triplicate wells and the resulting AP content in each well is 5.50 ng, 1.74 ng, 0.55 ng, 0.18 ng. 100 μl of pNNP reagent is then added to each of the standard wells. The plate is incubated at 37° C. for 4 h. A volume of 50 μl of 3M NaOH is added to all wells. The plate is read on a plate reader at 405 nm using the background subtraction option on blank wells filled with glycine buffer only. Additionally, the template is set up to indicate the concentration of AP-conjugate in each standard well [5.50 ng; 1.74 ng; 0.55 ng; 0.18 ng]. Results are indicated as amount of bound AP-conjugate in each sample.

Example 44 Alamar Blue Endothelial Cells Proliferation Assay

This assay may be used to quantitatively determine protein mediated inhibition of bFGF-induced proliferation of Bovine Lymphatic Endothelial Cells (LECs), Bovine Aortic Endothelial Cells (BAECs) or Human Microvascular Uterine Myometrial Cells (UTMECs). This assay incorporates a fluorometric growth indicator based on detection of metabolic activity. A standard Alamar Blue Proliferation Assay is prepared in EGM-2MV with 10 ng/ml of bFGF added as a source of endothelial cell stimulation. This assay may be used with a variety of endothelial cells with slight changes in growth medium and cell concentration. Dilutions of the protein batches to be tested are diluted as appropriate. Serum-free medium (GIBCO SFM) without bFGF is used as a non-stimulated control and Angiostatin or TSP-1 are included as a known inhibitory controls.

Briefly, LEC, BAECs or UTMECs are seeded in growth media at a density of 5000 to 2000 cells/well in a 96 well plate and placed at 37 degrees C. overnight. After the overnight incubation of the cells, the growth media is removed and replaced with GIBCO EC-SFM. The cells are treated with the appropriate dilutions of the protein of interest or control protein sample(s) (prepared in SFM) in triplicate wells with additional bPGF to a concentration of 10 ng/ml. Once the cells have been treated with the samples, the plate(s) is/are placed back in the 37° C. incubator for three days. After three days 10 ml of stock alamar blue (Biosource Cat# DAL100) is added to each well and the plate(s) is/are placed back in the 37° C. incubator for four hours. The plate(s) are then read at 530 nm excitation and 590 nm emission using the CytoFluor fluorescence reader. Direct output is recorded in relative fluorescence units.

Alamar blue is an oxidation-reduction indicator that both fluoresces and changes color in response to chemical reduction of growth medium resulting from cell growth. As cells grow in culture, innate metabolic activity results in a chemical reduction of the immediate surrounding environment. Reduction related to growth causes the indicator to change from oxidized (non-fluorescent blue) form to reduced (fluorescent red) form (i.e., stimulated proliferation will produce a stronger signal and inhibited proliferation will produce a weaker signal and the total signal is proportional to the total number of cells as well as their metabolic activity). The background level of activity is observed with the starvation medium alone. This is compared to the output observed from the positive control samples (bFGF in growth medium) and protein dilutions.

Example 45 Detection of Inhibition of a Mixed Lymphocyte Reaction

This assay can be used to detect and evaluate inhibition of a Mixed Lymphocyte Reaction (MLR) by gene products (e.g., isolated polypeptides). Inhibition of a MLR may be due to a direct effect on cell proliferation and viability, modulation of costimulatory molecules on interacting cells, modulation of adhesiveness between lymphocytes and accessory cells, or modulation of cytokine production by accessory cells. Multiple cells may be targeted by these polypeptides since the peripheral blood mononuclear fraction used in this assay includes T, B and natural killer lymphocytes, as well as monocytes and dendritic cells.

Polypeptides of interest found to inhibit the MLR may find application in diseases associated with lymphocyte and monocyte activation or proliferation. These include, but are not limited to, diseases such as asthma, arthritis, diabetes, inflammatory skin conditions, psoriasis, eczema, systemic lupus erythematosus, multiple sclerosis, glomerulonephritis, inflammatory bowel disease, crohn's disease, ulcerative colitis, arteriosclerosis, cirrhosis, graft vs. host disease, host vs. graft disease, hepatitis, leukemia and lymphoma.

Briefly, PBMCs from human donors are purified by density gradient centrifugation using Lymphocyte Separation Medium (LSM®, density 1.0770 g/ml, Organon Teknika Corporation, West Chester, Pa.). PBMCs from two donors are adjusted to 2×10⁶ cells/ml in RPMI-1640 (Life Technologies, Grand Island, N.Y.) supplemented with 10% FCS and 2 mM glutamine. PBMCs from a third donor is adjusted to 2×10⁵ cells/ml. Fifty microliters of PBMCs from each donor is added to wells of a 96-well round bottom microtiter plate. Dilutions of test materials (50 μl) is added in triplicate to microtiter wells. Test samples (of the protein of interest) are added for final dilution of 1:4; rhuIL-2 (R&D Systems, Minneapolis, Minn., catalog number 202-IL) is added to a final concentration of 1 μg/ml; anti-CD4 mAb (R&D Systems, clone 34930.11, catalog number MAB379) is added to a final concentration of 10 μg/ml. Cells are cultured for 7-8 days at 37° C. in 5% CO₂, and 1 μC of [³H] thymidine is added to wells for the last 16 hrs of culture. Cells are harvested and thymidine incorporation determined using a Packard TopCount. Data is expressed as the mean and standard deviation of triplicate determinations.

Samples of the protein of interest are screened in separate experiments and compared to the negative control treatment, anti-CD4 mAb, which inhibits proliferation of lymphocytes and the positive control treatment, IL-2 (either as recombinant material or supernatant), which enhances proliferation of lymphocytes.

One skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof.

Example 46 Assays for Protease Activity

The following assay may be used to assess protease activity of the polypeptides of the invention.

Gelatin and casein zymography are performed essentially as described (Heusen et al., Anal. Biochem., 102:196-202 (1980); Wilson et al., Journal of Urology, 149:653-658 (1993)). Samples are run on 10% polyacryamide/0.1% SDS gels containing 1% gelain orcasein, soaked in 2.5% triton at room temperature for 1 hour, and in 0.1M glycine, pH 8.3 at 37° C. 5 to 16 hours. After staining in amido black areas of proteolysis appear as clear areas against the blue-black background. Trypsin (Sigma T8642) is used as a positive control.

Protease activity is also determined by monitoring the cleavage of n-a-benzoyl-L-arginine ethyl ester (BAEE) (Sigma B-4500. Reactions are set up in (25 mMNaPO₄, 1 mM EDTA, and 1 mM BAEE), pH 7.5. Samples are added and the change in absorbance at 260 nm is monitored on the Beckman DU-6 spectrophotometer in the time-drive mode. Trypsin is used as a positive control.

Additional assays based upon the release of acid-soluble peptides from casein or hemoglobin measured as absorbance at 280 nm or colorimetrically using the Folin method are performed as described in Bergmeyer, et al., Methods of Enzymatic Analysis, 5 (1984). Other assays involve the solubilization of chromogenic substrates (Ward, Applied Science, 251-317 (1983)).

Example 47 Identifying Serine Protease Substrate Specificity

Methods known in the art or described herein may be used to determine the substrate specificity of the polypeptides of the present invention having serine protease activity. A preferred method of determining substrate specificity is by the use of positional scanning synthetic combinatorial libraries as described in GB 2 324 529 (incorporated herein in its entirety).

Example 48 Ligand Binding Assays

The following assay may be used to assess ligand binding activity of the polypeptides of the invention.

Ligand binding assays provide a direct method for ascertaining receptor pharmacology and are adaptable to a high throughput format. The purified ligand for a polypeptide is radiolabeled to high specific activity (50-2000 Ci/mmol) for binding studies. A determination is then made that the process of radiolabeling does not diminish the activity of the ligand towards its polypeptide. Assay conditions for buffers, ions, pH and other modulators such as nucleotides are optimized to establish a workable signal to noise ratio for both membrane and whole cell polypeptide sources. For these assays, specific polypeptide binding is defined as total associated radioactivity minus the radioactivity measured in the presence of an excess of unlabeled competing ligand. Where possible, more than one competing ligand is used to define residual nonspecific binding.

Example 49 Functional Assay in Xenopus Oocytes

Capped RNA transcripts from linearized plasmid templates encoding the polypeptides of the invention are synthesized in vitro with RNA polymerases in accordance with standard procedures. In vitro transcripts are suspended in water at a final concentration of 0.2 mg/ml. Ovarian lobes are removed from adult female toads, Stage V defolliculated oocytes are obtained, and RNA transcripts (10 ng/oocytc) are injected in a 50 nl bolus using a microinjection apparatus. Two electrode voltage clamps are used to measure the currents from individual Xenopus oocytes in response polypeptides and polypeptide agonist exposure. Recordings are made in Ca2+ free Barth's medium at room temperature. The Xenopus system can be used to screen known ligands and tissue/cell extracts for activating ligands.

Example 50 Microphysiometric Assays

Activation of a wide variety of secondary messenger systems results in extrusion of small amounts of acid from a cell. The acid formed is largely as a result of the increased metabolic activity required to fuel the intracellular signaling process. The pH changes in the media surrounding the cell are very small but are detectable by the CYTOSENSOR microphysiometer (Molecular Devices Ltd., Menlo Park, Calif.). The CYTOSENSOR is thus capable of detecting the activation of polypeptide which is coupled to an energy utilizing intracellular signaling pathway.

Example 51 Extract/Cell Supernatant Screening

A large number of mammalian receptors exist for which there remains, as yet, no cognate activating ligand (agonist). Thus, active ligands for these receptors may not be included within the ligands banks as identified to date. Accordingly, the polypeptides of the invention can also be functionally screened (using calcium, cAMP, microphysiometer, oocyte electrophysiology, etc., functional screens) against tissue extracts to identify its natural ligands. Extracts that produce positive functional responses can be sequentially subfractionated until an activating ligand is isolated and identified.

Example 52 Calcium and cAMP Functional Assays

Seven transmembrane receptors which are expressed in HEK 293 cells have been shown to be coupled functionally to activation of PLC and calcium mobilization and/or cAMP stimulation or inhibition. Basal calcium levels in the HEK 293 cells in receptor-transfected or vector control cells were observed to be in the normal, 100 nM to 200 nM, range. HEK 293 cells expressing recombinant receptors are loaded with fura 2 and in a single day >150 selected ligands or tissue/cell extracts are evaluated for agonist induced calcium mobilization. Similarly, HEK 293 cells expressing recombinant receptors are evaluated for the stimulation or inhibition of cAMP production using standard cAMP quantitation assays. Agonists presenting a calcium transient or cAMP fluctuation are tested in vector control cells to determine if the response is unique to the transfected cells expressing receptor.

Example 53 ATP-Binding Assay

The following assay may be used to assess ATP-binding activity of polypeptides of the invention.

ATP-binding activity of the polypeptides of the invention may be detected using the ATP-binding assay described in U.S. Pat. No. 5,858,719, which is herein incorporated by reference in its entirety. Briefly, ATP-binding to polypeptides of the invention is measured via photoaffinity labeling with 8-azido-ATP in a competition assay. Reaction mixtures containing 1 mg/ml of the ABC transport protein of the present invention are incubated with varying concentrations of ATP, or the non-hydrolyzable ATP analog adenyl-5′-imidodiphosphate for 10 minutes at 4° C. A mixture of 8-azido-ATP (Sigma Chem. Corp., St. Louis, Mo.) plus 8-azido-ATP (³²P-ATP) (5 mCi/μmol, ICN, Irvine Calif.) is added to a final concentration of 100 μM and 0.5 ml aliquots are placed in the wells of a porcelain spot plate on ice. The plate is irradiated using a short wave 254 nm UV lamp at a distance of 2.5 cm from the plate for two one-minute intervals with a one-minute cooling interval in between. The reaction is stopped by addition of dithiothreitol to a final concentration of 2 mM. The incubations are subjected to SDS-PAGE electrophoresis, dried, and autoradiographed. Protein bands corresponding to the particular polypeptides of the invention are excised, and the radioactivity quantified. A decrease in radioactivity with increasing ATP or adenyl-5′-imidodiphosphate provides a measure of ATP affinity to the polypeptides.

Example 54 Small Molecule Screening

This invention is particularly useful for screening therapeutic compounds by using the polypeptides of the invention, or binding fragments thereof, in any of a variety of drug screening techniques. The polypeptide or fragment employed in such a test may be affixed to a solid support, expressed on a cell surface, free in solution, or located intracellularly. One method of drug screening utilizes eukaryotic or prokaryotic host cells which are stably transformed with recombinant nucleic acids expressing the polypeptide or fragment. Drugs are screened against such transformed cells in competitive binding assays. One may measure, for example, the formulation of complexes between the agent being tested and polypeptide of the invention.

Thus, the present invention provides methods of screening for drugs or any other agents which affect activities mediated by the polypeptides of the invention. These methods comprise contacting such an agent with a polypeptide of the invention or fragment thereof and assaying for the presence of a complex between the agent and the polypeptide or fragment thereof, by methods well known in the art. In such a competitive binding assay, the agents to screen are typically labeled. Following incubation, free agent is separated from that present in bound form, and the amount of free or uncomplexed label is a measure of the ability of a particular agent to bind to the polypeptides of the invention.

Another technique for drug screening provides high throughput screening for compounds having suitable binding affinity to the polypeptides of the invention, and is described in great detail in European Patent Application 84/03564, published on Sep. 13, 1984, which is herein incorporated by reference in its entirety. Briefly stated, large numbers of different small molecule test compounds are synthesized on a solid substrate, such as plastic pins or some other surface. The test compounds are reacted with polypeptides of the invention and washed. Bound polypeptides are then detected by methods well known in the art. Purified polypeptides are coated directly onto plates for use in the aforementioned drug screening techniques. In addition, non-neutralizing antibodies may be used to capture the peptide and immobilize it on the solid support.

This invention also contemplates the use of competitive drug screening assays in which neutralizing antibodies capable of binding polypeptides of the invention specifically compete with a test compound for binding to the polypeptides or fragments thereof. In this manner, the antibodies are used to detect the presence of any peptide which shares one or more antigenic epitopes with a polypeptide of the invention.

Example 55 Phosphorylation Assay

In order to assay for phosphorylation activity of the polypeptides of the invention, a phosphorylation assay as described in U.S. Pat. No. 5,958,405 (which is herein incorporated by reference) is utilized. Briefly, phosphorylation activity may be measured by phosphorylation of a protein substrate using gamma-labeled ³²P-ATP and quantitation of the incorporated radioactivity using a gamma radioisotope counter. The polypeptides of the invention are incubated with the protein substrate, ³²P-ATP, and a kinase buffer. The ³²P incorporated into the substrate is then separated from free ³²P-ATP by electrophoresis, and the incorporated ³²P is counted and compared to a negative control. Radioactivity counts above the negative control are indicative of phosphorylation activity of the polypeptides of the invention.

Example 56 Detection of Phosphorylation Activity (Activation) of the Polypeptides of the Invention in the Presence of Polypeptide Ligands

Methods known in the art or described herein may be used to determine the phosphorylation activity of the polypeptides of the invention. A preferred method of determining phosphorylation activity is by the use of the tyrosine phosphorylation assay as described in U.S. Pat. No. 5,817,471 (incorporated herein by reference).

Example 57 Identification of Signal Transduction Proteins that Interact with Polypeptides of the Present Invention

The purified polypeptides of the invention are research tools for the identification, characterization and purification of additional signal transduction pathway proteins or receptor proteins. Briefly, labeled polypeptides of the invention are useful as reagents for the purification of molecules with which it interacts. In one embodiment of affinity purification, polypeptides of the invention are covalently coupled to a chromatography column. Cell-free extract derived from putative target cells, such as carcinoma tissues, is passed over the column, and molecules with appropriate affinity bind to the polypeptides of the invention. The protein complex is recovered from the column, dissociated, and the recovered molecule subjected to N-terminal protein sequencing. This amino acid sequence is then used to identify the captured molecule or to design degenerate oligonucleotide probes for cloning the relevant gene from an appropriate cDNA library.

Example 58 IL-6 Bioassay

To test the proliferative effects of the polypeptides of the invention, the IL-6 Bioassay as described by Marz et al. is utilized (Proc. Natl. Acad. Sci., U.S.A., 95:3251-56 (1998), which is herein incorporated by reference). Briefly, IL-6 dependent B9 murine cells are washed three times in IL-6 free medium and plated at a concentration of 5,000 cells per well in 50 μl, and 50 μl of the IL-6-like polypeptide is added. After 68 hrs. at 37° C., the number of viable cells is measured by adding the tetrazolium salt thiazolyl blue (MTT) and incubating for a further 4 hrs. at 37° C. B9 cells are lysed by SDS and optical density is measured at 570 nm. Controls containing IL-6 (positive) and no cytokine (negative) are utilized. Enhanced proliferation in the test sample(s) relative to the negative control is indicative of proliferative effects mediated by polypeptides of the invention.

Example 59 Support of Chicken Embryo Neuron Survival

To test whether sympathetic neuronal cell viability is supported by polypeptides of the invention, the chicken embryo neuronal survival assay of Senaldi et al is utilized (Proc. Natl. Acad. Sci., U.S.A., 96:11458-63 (1998), which is herein incorporated by reference). Briefly, motor and sympathetic neurons are isolated from chicken embryos, resuspended in L15 medium (with 10% FCS, glucose, sodium selenite, progesterone, conalbumin, putrescine, and insulin; Life Technologies, Rockville, Md.) and Dulbecco's modified Eagles medium [with 10% FCS, glutamine, penicillin, and 25 mM Hepes buffer (pH 7.2); Life Technologies, Rockville, Md.], respectively, and incubated at 37° C. in 5% CO₂ in the presence of different concentrations of the purified IL-6-like polypeptide, as well as a negative control lacking any cytokine. After 3 days, neuron survival is determined by evaluation of cellular morphology, and through the use of the colorimetric assay of Mosmann (Mosmann, T., J. Immunol. Methods, 65:55-63 (1983)). Enhanced neuronal cell viability as compared to the controls lacking cytokine is indicative of the ability of the inventive purified IL-6-like polypeptide(s) to enhance the survival of neuronal cells.

Example 60 Assay for Phosphatase Activity

The following assay may be used to assess serine/threonine phosphatase (PTPase) activity of the polypeptides of the invention.

In order to assay for serine/threonine phosphatase (PTPase) activity, assays can be utilized which are widely known to those skilled in the art. For example, the serine/threonine phosphatase (PSPase) activity is measured using a PSPase assay kit from New England Biolabs, Inc. Myelin basic protein (MyBP), a substrate for PSPase, is phosphorylated on serine and threonine residues with cAMP-dependent Protein Kinase in the presence of [³²P]ATP. Protein serine/threonine phosphatase activity is then determined by measuring the release of inorganic phosphate from 32P-labeled MyBP.

Example 61 Interaction of Serine/Threonine Phosphatases with Other Proteins

The polypeptides of the invention with serine/threonine phosphatase activity as determined in Example 60 are research tools for the identification, characterization and purification of additional interacting proteins or receptor proteins, or other signal transduction pathway proteins. Briefly, labeled polypeptide(s) of the invention is useful as a reagent for the purification of molecules with which it interacts. In one embodiment of affinity purification, polypeptide of the invention is covalently coupled to a chromatography column. Cell-free extract derived from putative target cells, such as neural or liver cells, is passed over the column, and molecules with appropriate affinity bind to the polypeptides of the invention. The polypeptides of the invention-complex is recovered from the column, dissociated, and the recovered molecule subjected to N-terminal protein sequencing. This amino acid sequence is then used to identify the captured molecule or to design degenerate oligonucleotide probes for cloning the relevant gene from an appropriate cDNA library.

Example 62 Assaying for Heparanase Activity

In order to assay for heparanase activity of the polypeptides of the invention, the heparanase assay described by Vlodavsky et al is utilized (Vlodavsky, I., et al., Nat. Med., 5:793-802 (1999)). Briefly, cell lysates, conditioned media or intact cells (1×10⁶ cells per 35-mm dish) are incubated for 18 hrs at 37° C., pH 6.2-6.6, with ³⁵S-labeled ECM or soluble ECM derived peak I proteoglycans. The incubation medium is centrifuged and the supernatant is analyzed by gel filtration on a Sepharose CL-6B column (0.9×30 cm). Fractions are eluted with PBS and their radioactivity is measured. Degradation fragments of heparan sulfate side chains are eluted from Sepharose 6B at 0.5<K_(av)<0.8 (peak II). Each experiment is done at least three times. Degradation fragments corresponding to “peak II,” as described by Vlodavsky et al., is indicative of the activity of the polypeptides of the invention in cleaving heparan sulfate.

Example 63 Immobilization of Biomolecules

This example provides a method for the stabilization of polypeptides of the invention in non-host cell lipid bilayer constructs (see, e.g., Bieri et al., Nature Biotech 17:1105-1108 (1999), hereby incorporated by reference in its entirety herein) which can be adapted for the study of polypeptides of the invention in the various functional assays described above. Briefly, carbohydrate-specific chemistry for biotinylation is used to confine a biotin tag to the extracellular domain of the polypeptides of the invention, thus allowing uniform orientation upon immobilization. A 50 uM solution of polypeptides of the invention in washed membranes is incubated with 20 mM NaIO4 and 1.5 mg/ml (4 mM) BACH or 2 mg/ml (7.5 mM) biotin-hydrazide for 1 hr at room temperature (reaction volume, 150 ul). Then the sample is dialyzed (Pierce Slidealizer Cassett, 10 kDa cutoff; Pierce Chemical Co., Rockford Ill.) at 4 C first for 5 h, exchanging the buffer after each hour, and finally for 12 h against 500 ml buffer R (0.15 M NaCl, 1 mM MgCl2, 10 mM sodium phosphate, pH7). Just before addition into a cuvette, the sample is diluted 1:5 in buffer ROG50 (Buffer R supplemented with 50 mM octylglucoside).

Example 64 TAQMAN

Quantitative PCR (QPCR). Total RNA from cells in culture are extracted by Trizol separation as recommended by the supplier (LifeTechnologies). (Total RNA is treated with DNase I (Life Technologies) to remove any contaminating genomic DNA before reverse transcription.) Total RNA (50 ng) is used in a one-step, 50 ul, RT-QPCR, consisting of Taqman Buffer A (Perkin-Elmer; 50 mM KCl/10 mM Tris, pH 8.3), 5.5 mM MgCl₂, 240 μM each dNTP, 0.4 units RNase inhibitor (Promega), 8% glycerol, 0.012% Tween-20, 0.05% gelatin, 0.3 uM primers, 0.1 uM probe, 0.025 units Amplitaq Gold (Perkin-Elmer) and 2.5 units Superscript II reverse transcriptase (Life Technologies). As a control for genomic contamination, parallel reactions are setup without reverse transcriptase. The relative abundance of (unknown) and 18S RNAs are assessed by using the Applied Biosystems Prism 7700 Sequence Detection System (Livak, K. J., Flood, S. J., Marmaro, J., Giusti, W. & Deetz, K. (1995) PCR Methods Appl. 4, 357-362). Reactions are carried out at 48° C. for 30 min, 95° C. for 10 min, followed by 40 cycles of 95° C. for 15 s, 60° C. for 1 min. Reactions are performed in triplicate.

Primers (f & r) and FRET probes sets are designed using Primer Express Software (Perkin-Elmer). Probes are labeled at the 5′-end with the reporter dye 6-FAM and on the 3′-end with the quencher dye TAMRA (Biosource International, Camarillo, Calif. or Perkin-Elmer).

Example 65 Assays for Metalloproteinase Activity

Metalloproteinases (EC 3.4.24.-) are peptide hydrolases which use metal ions, such as Zn²⁺, as the catalytic mechanism. Metalloproteinase activity of polypeptides of the present invention can be assayed according to the following methods.

Proteolysis of alpha-2-macroglobulin

To confirm protease activity, purified polypeptides of the invention are mixed with the substrate alpha-2-macroglobulin (0.2 unit/ml; Boehringer Mannheim, Germany) in 1× assay buffer (50 mM HEPES, pH 7.5, 0.2 M NaCl, 10 mM CaCl₂, 25 μM ZnCl₂ and 0.05% Brij-35) and incubated at 37° C. for 1-5 days. Trypsin is used as positive control. Negative controls contain only alpha-2-macroglobulin in assay buffer. The samples are collected and boiled in SDS-PAGE sample buffer containing 5% 2-mercaptoethanol for 5-min, then loaded onto 8% SDS-polyacrylamide gel. After electrophoresis the proteins are visualized by silver staining. Proteolysis is evident by the appearance of lower molecular weight bands as compared to the negative control.

Inhibition of alpha-2-macroglobulin proteolysis by inhibitors of metalloproteinases

Known metalloproteinase inhibitors (metal chelators (EDTA, EGTA, AND HgCl₂), peptide metalloproteinase inhibitors (TIMP-1 and TIMP-2), and commercial small molecule MMP inhibitors) are used to characterize the proteolytic activity of polypeptides of the invention. The three synthetic MMP inhibitors used are: MMP inhibitor I, [IC₅₀=1.0 μM against MMP-1 and MMP-8; IC₅₀=30 μM against MMP-9; IC₅₀=150 μM against MMP-3]; MMP-3 (stromelysin-1) inhibitor I [IC₅₀=5 μM against MMP-3], and MMP-3 inhibitor II [K_(i)=130 nM against MMP-3]; inhibitors available through Calbiochem, catalog # 444250, 444218, and 444225, respectively). Briefly, different concentrations of the small molecule MMP inhibitors are mixed with purified polypeptides of the invention (50 μg/ml) in 22.9 μl of 1×HEPES buffer (50 mM HEPES, pH 7.5, 0.2 M NaCl, 10 mM CaCl₂, 25 μM ZnCl₂ and 0.05% Brij-35) and incubated at room temperature (24° C.) for 2-hr, then 7.1 μl of substrate alpha-2-macroglobulin (0.2 unit/ml) is added and incubated at 37° C. for 20-hr. The reactions are stopped by adding 4× sample buffer and boiled immediately for 5 minutes. After SDS-PAGE, the protein bands are visualized by silver stain.

Synthetic Fluorogenic Peptide Substrates Cleavage Assay

The substrate specificity for polypeptides of the invention with demonstrated metalloproteinase activity can be determined using synthetic fluorogenic peptide substrates (purchased from BACHEM Bioscience Inc). Test substrates include, M-1985, M-2225, M-2105, M-2110, and M-2255. The first four are MMP substrates and the last one is a substrate of tumor necrosis factor-α (TNF-α) converting enzyme (TACE). All the substrates are prepared in 1:1 dimethyl sulfoxide (DMSO) and water. The stock solutions are 50-500 μM. Fluorescent assays are performed by using a Perkin Elmer LS 50B luminescence spectrometer equipped with a constant temperature water bath. The excitation λ is 328 nm and the emission λ is 393 nm. Briefly, the assay is carried out by incubating 176 μl 1×HEPES buffer (0.2 M NaCl, 10 mM CaCl₂, 0.05% Brij-35 and 50 mM HEPES, pH 7.5) with 4 μl of substrate solution (50 μM) at 25° C. for 15 minutes, and then adding 20 μl of a purified polypeptide of the invention into the assay cuvett. The final concentration of substrate is 1 μM. Initial hydrolysis rates are monitored for 30-min.

Example 66 Characterization of the cDNA Contained in a Deposited Plasmid

The size of the cDNA insert contained in a deposited plasmid may be routinely determined using techniques known in the art, such as PCR amplification using synthetic primers hybridizable to the 3′ and 5′ ends of the cDNA sequence. For example, two primers of 17-30 nucleotides derived from each end of the cDNA (i.e., hybridizable to the absolute 5′ nucleotide or the 3′ nucleotide end of the sequence of SEQ ID NO:X, respectively) are synthesized and used to amplify the cDNA using the deposited cDNA plasmid as a template. The polymerase chain reaction is carried out under routine conditions, for instance, in 25 ul of reaction mixture with 0.5 ug of the above cDNA template. A convenient reaction mixture is 1.5-5 mM MgCl₂, 0.01% (w/v) gelatin, 20 uM each of dATP, dCTP, dGTP, dTTP, 25 pmol of each primer and 0.25 Unit of Taq polymerase. Thirty five cycles of PCR (denaturation at 94 degree C. for 1 min; annealing at 55 degree C. for 1 min; elongation at 72 degree C. for 1 min) are performed with a Perkin-Elmer Cetus automated thermal cycler. The amplified product is analyzed by agarose gel electrophoresis. The PCR product is verified to be the selected sequence by subcloning and sequencing the DNA product. It will be clear that the invention may be practiced otherwise than as particularly described in the foregoing description and examples. Numerous modifications and variations of the present invention are possible in light of the above teachings and, therefore, are within the scope of the appended claims.

INCORPORATION BY REFERENCE

The entire disclosure of each document cited (including patents, patent applications, journal articles, abstracts, laboratory manuals, books, or other disclosures) in the Background of the Invention, Detailed Description, and Examples is hereby incorporated herein by reference. In addition, the sequence listing submitted herewith is incorporated herein by reference in its entirety. The specification and sequence listing of each of the following U.S. and PCT applications are herein incorporated by reference in their entirety (filing dates shown in format “year-month-day” (yyyy-mm-dd)): Application No. 60/278,650 filed on 2001 Mar. 27, application Ser. No. 09/950,082 filed on 2001 Sep. 12, application Ser. No. 09/950,083 filed on 2001 Sep. 12, Application No. 60/306,171 filed on 19 Jul. 2001, application Ser. No. 09/833,245 filed on 2001 Apr. 12, Application No. PCT/US01/11988 filed on 2001 Apr. 12, Application No. 60/331,287 filed on 2001 Nov. 13, Application No. 60/277,340 filed on 2001 Mar. 21, Application No. PCT/US00/06043 filed on 2000 Mar. 9, Application No. PCT/US00/06012 filed on 2000 Mar. 9, Application No. PCT/US00/06058 filed on 2000 Mar. 9, Application No. PCT/US00/06044 filed on 2000 Mar. 9, Application No. PCT/US00/06059 filed on 2000 Mar. 9, Application No. PCT/US00/06042 filed on 2000 Mar. 9, Application No. PCT/US00/06014 filed on 2000 Mar. 9, Application No. PCT/US00/06013 filed on 2000 Mar. 9, Application No. PCT/US00/06049 filed on 2000 Mar. 9, Application No. PCT/US00/06057 filed on 2000 Mar. 9, Application No. PCT/US00/06824 filed on 2000 Mar. 16, Application No. PCT/US00/06765 filed on 2000 Mar. 16, Application No. PCT/US00/06792 filed on 2000 Mar. 16, Application No. PCT/US00/06830 filed on 2000 Mar. 16, Application No. PCT/US00/06782 filed on 2000 Mar. 16, 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PCT/US00/07724 filed on 2000 Mar. 23, Application No. PCT/US00/14929 filed on 2000 Jun. 1, Application No. PCT/US00/07722 filed on 2000 Mar. 23, Application No. PCT/US00/07578 filed on 2000 Mar. 23, Application No. PCT/US00/07726 filed on 2000 Mar. 23, Application No. PCT/US00/07677 filed on 2000 Mar. 23, Application No. PCT/US00/07725 filed on 2000 Mar. 23, Application No. PCT/US00/09070 filed on 2000 Apr. 6, Application No. PCT/US00/08982 filed on 2000 Apr. 6, Application No. PCT/US00/08983 filed on 2000 Apr. 6, Application No. PCT/US00/09067 filed on 2000 Apr. 6, Application No. PCT/US00/09066 filed on 2000 Apr. 6, Application No. PCT/US00/09068 filed on 2000 Apr. 6, Application No. PCT/US00/08981 filed on 2000 Apr. 6, Application No. PCT/US00/08980 filed on 2000 Apr. 6, Application No. PCT/US00/09071 filed on 2000 Apr. 6, Application No. PCT/US00/09069 filed on 2000 Apr. 6, Application No. PCT/US00/15136 filed on 2000 Jun. 1, Application No. PCT/US00/14926 filed on 2000 Jun. 1, Application No. PCT/US00/14963 filed on 2000 Jun. 1, Application No. PCT/US00/15135 filed on 2000 Jun. 1, Application No. PCT/US00/14934 filed on 2000 Jun. 1, Application No. PCT/US00/14933 filed on 2000 Jun. 1, Application No. PCT/US00/15137 filed on 2000 Jun. 1, Application No. PCT/US00/14928 filed on 2000 Jun. 1, Application No. PCT/US00/14973 filed on 2000 Jun. 1, Application No. PCT/US00/14964 filed on 2000 Jun. 1, Application No. PCT/US00/26376 filed on 2000 Sep. 26, Application No. PCT/US00/26371 filed on 2000 Sep. 26, Application No. PCT/US00/26324 filed on 2000 Sep. 26, Application No. PCT/US00/26323 filed on 2000 Sep. 26, Application No. PCT/US00/26337 filed on 2000 Sep. 26, Application No. PCT/US01/13318 filed on 2001 Apr. 27, Application No. U.S. 60/124,146 filed on 1999 Mar. 12, Application No. U.S. 60/167,061 filed on 1999 Nov. 23, Application No. U.S. 60/124,093 filed on 1999 Mar. 12, Application No. U.S. 60/166,989 filed on 1999 Nov. 23, Application No. U.S. 60/124,145 filed on 1999 Mar. 12, Application No. U.S. 60/168,654 filed on 1999 Dec. 3, Application No. U.S. 60/124,099 filed on 1999 Mar. 12, Application No. U.S. 60/168,661 filed on 1999 Dec. 3, Application No. U.S. 60/124,096 filed on 1999 Mar. 12, Application No. U.S. 60/168,622 filed on 1999 Dec. 3, Application No. U.S. 60/124,143 filed on 1999 Mar. 12, Application No. U.S. 60/168,663 filed on 1999 Dec. 3, Application No. U.S. 60/124,095 filed on 1999 Mar. 12, Application No. U.S. 60/138,598 filed on 1999 Jun. 11, Application No. U.S. 60/168,665 filed on 1999 Dec. 3, Application No. U.S. 60/125,360 filed on 1999 Mar. 19, Application No. U.S. 60/138,626 filed on 1999 Jun. 11, Application No. U.S. 60/168,662 filed on 1999 Dec. 3, Application No. U.S. 60/124,144 filed on 1999 Mar. 12, Application No. U.S. 60/138,574 filed on 1999 Jun. 11, Application No. U.S. 60/168,667 filed on 1999 Dec. 3, Application No. U.S. 60/124,142 filed on 1999 Mar. 12, Application No. U.S. 60/138,597 filed on 1999 Jun. 11, Application No. U.S. 60/168,666 filed on 1999 Dec. 3, Application No. U.S. 60/125,359 filed on 1999 Mar. 19, Application No. U.S. 60/168,664 filed on 1999 Dec. 3, Application No. U.S. 60/126,051 filed on 1999 Mar. 23, Application No. U.S. 60/169,906 filed on 1999 Dec. 10, Application No. U.S. 60/125,362 filed on 1999 Mar. 19, Application No. U.S. 60/169,980 filed on 1999 Dec. 10, Application No. U.S. 60/125,361 filed on 1999 Mar. 19, Application No. U.S. 60/169,910 filed on 1999 Dec. 10, Application No. U.S. 60/125,812 filed on 1999 Mar. 23, Application No. U.S. 60/169,936 filed on 1999 Dec. 10, Application No. U.S. 60/126,054 filed on 1999 Mar. 23, Application No. U.S. 60/169,916 filed on 1999 Dec. 10, Application No. U.S. 60/125,815 filed on 1999 Mar. 23, Application No. U.S. 60/169,946 filed on 1999 Dec. 10, Application No. U.S. 60/125,358 filed on 1999 Mar. 19, Application No. U.S. 60/169,616 filed on 1999 Dec. 8, Application No. U.S. 60/125,364 filed on 1999 Mar. 19, Application No. U.S. 60/169,623 filed on 1999 Dec. 8, Application No. U.S. 60/125,363 filed on 1999 Mar. 19, Application No. U.S. 60/169,617 filed on 1999 Dec. 8, Application No. U.S. 60/126,502 filed on 1999 Mar. 26, Application No. U.S. 60/172,410 filed on 1999 Dec. 17, Application No. U.S. 60/126,503 filed on 1999 Mar. 26, Application No. U.S. 60/172,409 filed on 1999 Dec. 17, Application No. U.S. 60/126,505 filed on 1999 Mar. 26, Application No. U.S. 60/172,412 filed on 1999 Dec. 17, Application No. U.S. 60/126,594 filed on 1999 Mar. 26, Application No. U.S. 60/172,408 filed on 1999 Dec. 17, Application No. U.S. 60/126,511 filed on 1999 Mar. 26, Application No. U.S. 60/172,413 filed on 1999 Dec. 17, Application No. U.S. 60/126,595 filed on 1999 Mar. 26, Application No. U.S. 60/171,549 filed on 1999 Dec. 22, Application No. U.S. 60/126,598 filed on 1999 Mar. 26, Application No. U.S. 60/171,504 filed on 1999 Dec. 22, Application No. U.S. 60/126,596 filed on 1999 Mar. 26, Application No. U.S. 60/171,552 filed on 1999 Dec. 22, Application No. U.S. 60/126,600 filed on 1999 Mar. 26, Application No. U.S. 60/171,550 filed on 1999 Dec. 22, Application No. U.S. 60/126,501 filed on 1999 Mar. 26, Application No. U.S. 60/171,551 filed on 1999 Dec. 22, Application No. U.S. 60/126,504 filed on 1999 Mar. 26, Application No. U.S. 60/174,847 filed on 2000 Jan. 7, Application No. U.S. 60/126,509 filed on 1999 Mar. 26, Application No. U.S. 60/174,853 filed on 2000 Jan. 7, Application No. U.S. 60/126,506 filed on 1999 Mar. 26, Application No. U.S. 60/174,852 filed on 2000 Jan. 7, Application No. U.S. 60/242,710 filed on 2000 Oct. 25, Application No. U.S. 60/126,510 filed on 1999 Mar. 26, Application No. U.S. 60/174,850 filed on 2000 Jan. 7, Application No. U.S. 60/138,573 filed on 1999 Jun. 11, Application No. U.S. 60/174,851 filed on 2000 Jan. 7, Application No. U.S. 60/126,508 filed on 1999 Mar. 26, Application No. U.S. 60/174,871 filed on 2000 Jan. 7, Application No. U.S. 60/126,507 filed on 1999 Mar. 26, Application No. U.S. 60/174,872 filed on 2000 Jan. 7, Application No. U.S. 60/126,597 filed on 1999 Mar. 26, Application No. U.S. 60/174,877 filed on 2000 Jan. 7, Application No. U.S. 60/126,601 filed on 1999 Mar. 26, Application No. U.S. 60/154,373 filed on 1999 Sep. 17, Application No. U.S. 60/176,064 filed on 2000 Jan. 14, Application No. U.S. 60/126,602 filed on 1999 Mar. 26, Application No. U.S. 60/176,063 filed on 2000 Jan. 14, Application No. U.S. 60/128,695 filed on 1999 Apr. 9, Application No. U.S. 60/176,052 filed on 2000 Jan. 14, Application No. U.S. 60/128,696 filed on 1999 Apr. 9, Application No. U.S. 60/176,069 filed on 2000 Jan. 14, Application No. U.S. 60/128,703 filed on 1999 Apr. 9, Application No. U.S. 60/176,068 filed on 2000 Jan. 14, Application No. U.S. 60/128,697 filed on 1999 Apr. 9, Application No. U.S. 60/176,929 filed on 2000 Jan. 20, Application No. U.S. 60/128,698 filed on 1999 Apr. 9, Application No. U.S. 60/176,926 filed on 2000 Jan. 20, Application No. U.S. 60/128,699 filed on 1999 Apr. 9, Application No. U.S. 60/177,050 filed on 2000 Jan. 20, Application No. U.S. 60/128,701 filed on 1999 Apr. 9, Application No. U.S. 60/177,166 filed on 2000 Jan. 20, Application No. U.S. 60/128,700 filed on 1999 Apr. 9, Application No. U.S. 60/176,930 filed on 2000 Jan. 20, Application No. U.S. 60/128,694 filed on 1999 Apr. 9, Application No. U.S. 60/176,931 filed on 2000 Jan. 20, Application No. U.S. 60/128,702 filed on 1999 Apr. 9, Application No. U.S. 60/177,049 filed on 2000 Jan. 20, Application No. U.S. 60/138,629 filed on 1999 Jun. 11, Application No. U.S. 60/138,628 filed on 1999 Jun. 11, Application No. U.S. 60/138,631 filed on 1999 Jun. 11, Application No. U.S. 60/138,632 filed on 1999 Jun. 11, Application No. U.S. 60/138,599 filed on 1999 Jun. 11, Application No. U.S. 60/138,572 filed on 1999 Jun. 11, Application No. U.S. 60/138,625 filed on 1999 Jun. 11, Application No. U.S. 60/138,633 filed on 1999 Jun. 11, Application No. U.S. 60/138,630 filed on 1999 Jun. 11, Application No. U.S. 60/138,627 filed on 1999 Jun. 11, Application No. U.S. 60/155,808 filed on 1999 Sep. 27, Application No. U.S. 60/155,804 filed on 1999 Sep. 27, Application No. U.S. 60/155,807 filed on 1999 Sep. 27, Application No. U.S. 60/155,805 filed on 1999 Sep. 27, Application No. U.S. 60/155,806 filed on 1999 Sep. 27, Application No. U.S. 60/201,194 filed on 2000 May 2, and Application No. U.S. 60/212,142 filed on 2000 Jun. 16. LENGTHY TABLE The patent application contains a lengthy table section. A copy of the table is available in electronic form from the USPTO web site (http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20070032413A1) An electronic copy of the table will also be available from the USPTO upon request and payment of the fee set forth in 37 CFR 1.19(b)(3). 

1-32. (canceled)
 33. An isolated polypeptide comprising amino acid residues 29 to 139 of SEQ ID NO:3118.
 34. The isolated polypeptide of claim 33 which comprises amino acid residues 2 to 139 of SEQ ID NO:3118.
 35. The isolated polypeptide of claim 33 which comprises amino acid residues 1 to 139 of SEQ ID NO:3118.
 36. The polypeptide of claim 33 which further comprises a heterologous polypeptide sequence.
 37. A composition comprising the polypeptide of claim 33 and a carrier.
 38. An isolated protein produced by the method comprising: (a) expressing the polypeptide of claim 33 by a cell; and (b) recovering said protein.
 39. An isolated polypeptide comprising the amino acid sequence of the secreted portion of the polypeptide encoded by the HADDE71 cDNA contained in ATCC Deposit No.
 203917. 40. The isolated polypeptide of claim 39 which comprises the amino acid sequence of the complete polypeptide encoded by the HADDE71 cDNA contained in ATCC Deposit No. 203917, excepting the N-terminal methionine.
 41. The isolated polypeptide of claim 39 which comprises the amino acid sequence of the complete polypeptide encoded by the HADDE71 cDNA contained in ATCC Deposit No.
 203917. 42. The polypeptide of claim 39 which further comprises a heterologous polypeptide sequence.
 43. A composition comprising the polypeptide of claim 39 and a carrier.
 44. An isolated protein produced by the method comprising: (a) expressing the protein of claim 39 by a cell; and (b) recovering said protein. 45-54. (canceled)
 55. An isolated polypeptide consisting of at least 30 contiguous amino acid residues of amino acid residues 1 to 139 of SEQ ID:3118, wherein said polypeptide comprises amino acid residues 1-20, 21-40, 41-60, 61-80, or 81-100 of SEQ ID:3118.
 56. The polypeptide of claim 55 which further comprises a heterologous polypeptide sequence.
 57. A composition comprising the polypeptide of claim 55 and a carrier.
 58. An isolated protein produced by the method comprising: (a) expressing the polypeptide of claim 55 by a cell; and (b) recovering said protein.
 59. An isolated polypeptide consisting of at least 30 contiguous amino acid residues of the complete polypeptide encoded by the HADDE71 cDNA clone in ATCC Deposit No. 203917, wherein said polypeptide comprises amino acid residues 1-20, 21-40, 41-60, 61-80, or 81-100 of the HADDE71 cDNA clone in ATCC Deposit No.
 203917. 60. The polypeptide of claim 59 which further comprises a heterologous polypeptide sequence.
 61. A composition comprising the polypeptide of claim 59 and a carrier.
 62. An isolated protein produced by the method comprising: (a) expressing the polypeptide of claim 59 by a cell; and (b) recovering said protein. 